KR101820515B1

KR101820515B1 - A composition for preventing or inhibiting a metastasis of a primary thyroid cancer

Info

Publication number: KR101820515B1
Application number: KR1020150010142A
Authority: KR
Inventors: 김원배; 김원구; 고정민; 전민지
Original assignee: 재단법인 아산사회복지재단
Priority date: 2015-01-21
Filing date: 2015-01-21
Publication date: 2018-01-22
Also published as: KR20160090172A

Abstract

본 발명은 원발성 갑상선암 전이 예방 또는 억제용 약제학적 조성물에 관한 것이다. 본 발명자들은 Slit-Robo 경로의 활성 저하는 큰 종양 크기, 림프절 전이, 원격전이와 재발 등의 갑상선암의 나쁜 예후와 연관 있음을 최초로 규명하였다. 이에 본 발명자들은 인간 SLIT2 (Slit homolog 2) 단백질 또는 유전자를 이용하여 원발성 갑상선암 전이를 억제할 수 있는 치료제를 제안한다. 본 발명은 최초로 갑상선암에서 종양 억제 인자로서의 Slit-Robo 경로의 역할을 확인하였으며, 새로운 바이오마커 및 새로운 분자 치료 표적으로 Slit2의 가능성을 제시한다.The present invention relates to a pharmaceutical composition for preventing or inhibiting primary thyroid cancer metastasis. The inventors of the present invention have for the first time determined that the deactivation of the Slit-Robo pathway is associated with a bad prognosis of thyroid cancer including large tumor size, lymph node metastasis, distant metastasis and recurrence. Accordingly, the present inventors propose a therapeutic agent capable of inhibiting primary thyroid cancer metastasis using human SLIT2 (Slit homolog 2) protein or gene. The present invention first confirmed the role of the Slit-Robo pathway as a tumor suppressor in thyroid cancer and suggests the possibility of Slit2 as a new biomarker and a new molecular therapy target.

Description

[0001] The present invention relates to a composition for preventing or inhibiting metastasis of primary thyroid cancer,

본 발명은 원발성 갑상선암 전이 예방 또는 억제용 조성물에 관한 것이다.
The present invention relates to a composition for preventing or inhibiting primary thyroid cancer metastasis.

전체 갑상선암의 90%를 차지하는 분화 갑상선암은 유두암과 여포암을 총칭한다. 최근 수 십여 년간 분화 갑상선암의 발생은 전세계적으로 증가하였지만 대부분 적절한 수술적 치료와 방사성 요오드 동위원소 치료로 양호한 예후를 보여 10년 생존율이 90%에 이른다.¹⁾ 수술과 방사성 요오드 동위원소를 포함하는 초치료가 끝나고 나면 환자들은 갑상선 호르몬 보충 요법과 함께 주기적으로 혈청 내 티로글로불린(thyroglobulin)을 측정하고 경부 초음파 등의 검사를 시행하여 재발 여부를 확인한다. 약 30%의 환자는 재발하며, 국소 재발의 경우 재수술 등의 치료를 받게된다. 약 15%의 환자에서는 수술 전 및 수술 후 원격 전이가 발견되는데, 원격전이의 절반가량은 폐에서 발생하고 그 다음으로는 뼈가 흔하다. 원격전이가 발견되면 갑상선암으로 인한 사망이 급격히 증가하는데, 이전 연구는 10년 생존율이 약 40% 라고 보고하였다.^2,3) 반복적인 국소 재발 또는 원격전이를 동반한 진행성 고위험 갑상선암 환자들의 경우 수술이 불가능하다면 방사성 요오드 치료가 유일한 치료이나 약 30%의 환자들만 방사성 요오드 치료에 반응을 보여 보다 효과적인 치료방법이 필요하다. ^2,4) Differential thyroid cancer, which accounts for 90% of all thyroid cancer, is called papillary cancer and follicular cancer. In recent decades, the incidence of differentiated thyroid cancer has increased worldwide, but most of them have good prognosis due to appropriate surgical treatment and radioiodine isotope therapy, and the 10-year survival rate reaches 90%. ^After surgery and radioactive iodine ischemic therapy, the patients are followed up periodically with thyroid hormone replacement therapy, and serum thyroglobulin is measured, and a cervical ultrasonogram is performed to confirm the recurrence. Approximately 30% of patients are relapsed, and in the case of local recurrence, they are treated for reoperation. In about 15% of patients, preoperative and postoperative distant metastases are found, with about half of the distant metastases occurring in the lungs, followed by bone. When remote metastases are found, death from thyroid cancer is dramatically increased. Previous studies reported a 10-year survival rate of about 40%. ^{2,3) In} patients with advanced high-risk thyroid carcinoma with recurrent local recurrence or distant metastasis, radioiodine therapy is the only treatment or only about 30% of patients respond to radioactive iodine treatment if surgery is not possible. Do. ^2,4)

최근 10여 년 사이 갑상선암의 발생과 관련된 세포 신호 전달체계와 유전자 이상에 대한 연구가 활발히 진행되었다.⁵⁾ 갑상선암에서 가장 잘 알려진 것은 MAPK (mitogen-activated protein kinase)와 PI3K (phosphatidyl inositol 3-kinase)/AKT(v-akt murine thymoma viral oncogene) 경로로서 이들 중 BRAF, RAS, RET/PTC, PAX8/PPARG 돌연변이가 가장 잘 알려진 주된 유전자 이상들이다.⁶⁾ 갑상선 유두암에서 가장 흔한 유전자 이상은 BRAF 점돌연변이로 600번 아미노산인 발린이 글루탐산으로 치환되는 것이 대표적이며(V600E) 외국의 경우 30-40%, 우리나라에서는 60-80%의 유두암이 BRAF 변이에 의해 발생하는 것으로 알려졌다. 여포암에서는 RAS 돌연변이가 가장 흔히 발견되고(20-35%) NRAS 61코돈과 HRAS의 61코돈의 돌연변이가 대표적으로, 원격전이의 유발과 연관이 있을 것으로 알려졌다.⁷⁾ 그 외 RET/PTC 재배열은 방사선 노출에 의한 유두암 발생에 관련된 것으로 생각되며 약 15-40%에서 발견되고, PAX8/PPARG 재배열은 여포암의 30-40%에서 관찰되는 것으로 알려졌다. 갑상선암은 또한 혈관이 매우 발달하는 종양으로 VEGF (Vascular Endotehlial Growth Factor) 및 VEGFR(VEGF receptors) 의 과발현도 흔히 관찰된다.^4,5) In recent decades, studies on cell signal transduction systems and gene abnormalities related to the development of thyroid cancer have been actively conducted. ^{5) The} most well-known pathway for thyroid cancer is mitogen-activated protein kinase (MAPK) and phosphatidyl inositol 3-kinase / AKT (v-akt murine thymoma viral oncogene) pathways. Among these, BRAF, RAS, RET / PTC, PAX8 / The PPARG mutation is the most well known major gene abnormality. ^{6) The} most common gene abnormality in thyroid papillary cancer is BRAF point mutation, which is a substitution of glutamate (V600E), which is a 600 amino acid amino acid, in 30-40% of foreign countries and 60-80% of papillary cancer in Korea is BRAF mutation . Ras mutations are most commonly found in follicular carcinomas (20-35%) and mutations in the 61 codons of the NRAS 61 codon and HRAS are typically associated with the induction of distant metastasis. ⁷⁾ Other RET / PTC rearrangements are believed to be associated with the development of papillary carcinoma by radiation exposure, and are found in approximately 15-40%, and PAX8 / PPARG rearrangements are observed in 30-40% of follicular cancers. Thyroid carcinoma is also a highly vascular tumor and overexpression of VEGF (VEGF) and VEGFR (VEGF receptors) is frequently observed. ^4,5)

갑상선암의 발생과 진행에 관한 분자생물학적 과정에 대한 이해의 진전으로, 이를 바탕으로 한 새로운 진단 및 예후의 바이오마커에 대한 연구가 진행되고 있다. 가장 많은 연구가 이루어진 것은 역시 BRAF V600E 돌연변이에 관한 것으로 이는 림프절 전이와 피막 침범, 재발 등의 진행성 갑상선암의 특성과 연관이 있으며 실험실적 연구에서 BRAF V600E 돌연변이를 유발하면 VEGF와 MET 등의 과발현을 유도하여 종양의 공격성을 유발한다. 또한 BRAF V600E돌연변이는 갑상선 세포로의 요오드 섭취를 담당하는 Sodium-Iodide Sympoter (NIS) 유전자의 발현을 억제하여 방사성 동위원소 불응성 유발에 기여하는 것으로 보고되었다.^8,9) 때문에 BRAF V600E 돌연변이를 진단과 예후 예측의 바이오마커로 이용하려는 시도들이 있는데, 한 연구에서 BRAF V600E 돌연변이를 진단 검체에서 확인하는 것이 갑상선암 예측에 매우 높은 특이도와 양성 예측도를 보였다.¹⁰⁾ The development of understanding of molecular biology of thyroid carcinogenesis and progression has led to the development of new biomarkers for diagnosis and prognosis. The BRAF V600E mutation is associated with the characteristics of advanced thyroid cancer such as lymph node metastasis, invasion of the capsule, and recurrence. Experimental studies have shown that overexpression of VEGF and MET is induced by mutation of BRAF V600E Causing aggression of the tumor. In addition, the BRAF V600E mutation has been reported to inhibit the expression of the sodium-iodide sympoter (NIS) gene, which is responsible for iodine uptake into thyroid cells, contributing to the induction of radioisotope rejection. ^8,9), because there are attempts to use the BRAF V600E mutation as a biomarker for diagnosis and prognosis, it was a very high positive predictive value and specificity in predicting thyroid cancer to determine the BRAF V600E mutation in diagnostic specimens from the study. ¹⁰⁾

또한 진행성, 방사성 요오드 동위원소 불응성 갑상선암에 대해 많은 인산화효소억제제가 분자 표적 치료제로서 이용되고 있다. 그 중 대표적으로, sorafenib은 RET, BRAF 등의 MAPK 경로와 VEGFR, PDGFR (platelet-derived growth factor receptor) 을 억제하며 3상 임상연구에서 의미 있는 무진행 생존기간(progression-free survival) 연장 효과가 확인되었다.¹¹⁾ 또 다른 연구에서는 MEK 억제제인 selumetinib이 방사성 요오드 동위원소 불응성 갑상선암의 요오드 섭취능을 충분히 회복시킴을 보고하였다.¹²⁾ 이 외에도 많은 약제에 대한 연구가 진행 중이지만 일부 환자에서만 치료에 대한 반응을 보이고 부작용으로 인해 치료를 중단해야 하는 경우도 빈번하며, 대다수의 환자에서 경과 호전 후 다시 병이 진행하게 되는 등 한계가 있어 더 효과적인 표적치료제에 대한 필요성이 상존한다.¹³⁾ In addition, a number of phosphorylase inhibitors for progressive, radioactive iodine-deficient thyroid cancer have been used as molecular target therapeutics. For example, sorafenib inhibits MAPK pathways such as RET and BRAF, VEGFR and platelet-derived growth factor receptor (PDGFR), and a significant prolongation of progression-free survival . ¹¹⁾ In another study, it was reported that the MEK inhibitor, selumetinib, fully restored iodine uptake of radioactive iodine-deficient thyroid cancer. ¹²⁾ In addition, many medications are being studied, but some patients respond to treatment only, and treatment is often stopped due to side effects. In most patients, the disease progresses after the progress of the disease. There is a need for more effective targeted therapies. ¹³⁾

최근 다른 암종 연구에서 액손 유도 분자들이 새로운 분자생물학적 표적으로 대두되고 있다.¹⁴⁾ Slit-Robo 경로는 그 중 하나인데, Slit 당단백은 대표적인 액손 유도 분자로 다양한 조직과 세포에서 분비되며 생리학적 활성은 그 수용체인 Roundabout (Robo)을 통한다. 1800년대 후반, 액손 유도 분자가 존재한다는 것이 처음 알려진 이 후 많은 연구가 이루어져왔는데 이들은 처음에는 신경계의 발생과정 동안에 뉴런과 액손의 이동을 조절하는 것으로만 생각되었지만 점차 심장, 폐, 신장, 유방 조직 등 다양한 조직에서 다양한 세포 내 과정을 조절하는 인자로서의 역할이 있는 것으로 알려졌으며 특히 최근에는 세포의 성장과 부착능, 이동능의 조절에 중요한 역할이 있음이 밝혀져 다양한 암의 발생 및 진행과정에 영향을 미치는 것으로 생각되고 있다.¹⁵⁾ Recently, axon-derived molecules have emerged as new molecular biologic targets in other carcinogenesis studies. ¹⁴⁾ Slit-Robo pathway is one of them. Slit glycoprotein is a typical axon-inducing molecule secreted from various tissues and cells. Its physiological activity is through its receptor, Roundabout (Robo). In the late 1800s, a number of studies have been reported since the first known existence of axons-inducing molecules, which initially seemed to modulate the movement of neurons and axons during the development of the nervous system, but gradually increased in the heart, lung, It has been known that it plays a role as a factor regulating various intracellular processes in various tissues. In recent years, it has been shown that it plays an important role in regulation of cell growth, adhesion, and migration, . ¹⁵⁾

Slit은 인체에서 Slit 1-3의 세 가지 형태로 존재하는데, 그 중 가장 많이 알려진 것이 Slit2로서 정상 조직과 비교하여 신장암¹⁶ ⁾, 대장암¹⁷ ⁾, 폐암 및 유방암¹⁸ ⁾ 등에서는 Slit2의 발현이 감소되었음이 확인된 바 있고, Slit2를 knock-down하였을 때 위암의 성장과 전이능이 활성화 되고¹⁹ ⁾ 최근의 한 연구는 Slit2-Robo 경로가 억제되었을 때 췌장암의 신경 침습과 전이가 증가함을 발표하였다.²⁰⁾ Slit1의 역할에 대해서는 아직 알려진 것이 없고 최근 Slit3는 배아 혈관 신생에 역할이 있는 것으로 알려졌다. Robo 또한 인체에서 Robo 1-4의 네 가지 형태로 존재하는데 Robo 1,2,3은 비슷한 구조와 기능을 가지고 있으나 Robo 4는 상당히 다른 구조와 기능을 가지고 있으며 특히 혈관 신생에 중요한 역할을 할 것으로 알려졌다.^21,22) 최근 한 연구에서는 췌장암에서 Robo 2 발현이 억제되면 불량한 생존율을 보임을 발표하기도 하였다.²³⁾ The Slit is to exist in the human body in three forms of Slit 1-3, etc. As it Slit2 best known of which renal cell ^{carcinoma. ¹⁶⁾} as compared to normal tissue, cancer ^{^17),} lung cancer and breast cancer ^¹⁸⁾ Expression of Slit2 and the check is reduced under the bar, when the Slit2 hayeoteul knock-down and the activation of gastric cancer growth and metastasis ability ^{^19),} a recent research has announced that the nerve invasion and metastasis of pancreatic cancer increases when the Slit2-Robo path is suppressed . ²⁰⁾ There is no known role for Slit1, and Slit3 has recently been shown to play a role in embryonic angiogenesis. Robo is also present in the human body in four forms of Robo 1-4. Robo 1, 2 and 3 have similar structures and functions, but Robo 4 has a significantly different structure and function, and plays an important role in angiogenesis . ^{21, 22). One} recent study has reported that poor survival of pancreatic cancer is inhibited by inhibition of Robo2 expression. ²³⁾

Slit과 Robo 아형의 각각의 역할과 서로 다른 아형과의 상호작용이 어떻게 다른 역할을 나타내는지에 대해서는 확실하지 않다. 현재까지 가장 많이 연구된 것은 Slit2와 Robo1의 상호작용으로서 유방암 및 폐암 유래 세포주를 이용한 연구에서 이들의 결합은 PI3K/Akt 경로 억제와 glycogen synthase kinase-3 beta (GSK-3b)의 활성을 통해 beta-catenin의 핵 내 이동은 억제하고 E-cadherin의 안정화를 유도하여 세포 간의 연결을 안정화하고 세포 이동과 성장을 억제하였다.^24,25) 또한 교모세포종 유래 세포를 이용한 연구에서Slit2와 Robo1의 결합은 Rho GTPase를 통해 Cdc42 등 액틴 세포골격 조절 인자들을 억제함으로써 세포 이동능을 억제하였다²⁶ ⁾ (도 1). 하지만 대장암 유래 세포주를 이용한 한 연구에서는Slit2-Robo1의 발현 증가가 유비퀴틴접합단백질의 활성을 유도하여 오히려 E-cadherin의 분해를 촉진하고 세포 성장과 상피-간엽 전환(epithelial-mesenchmal transition)을 촉진함을 보고하여 전혀 반대의 결과를 보였다.²⁷⁾ 이에 Slit과 Robo경로는 암세포 종류와 그 단계에 따라 역할이 상이할 것으로 생각되고 있다.¹⁵⁾ It is unclear how the role of each of the Slit and Robo subtypes and their different subtypes play a different role. Most of the studies that have been conducted so far have focused on the interaction between Slit2 and Robo1 in breast cancer and lung cancer-derived cell lines, and their binding has been confirmed by the inhibition of PI3K / Akt pathway and the activation of glycogen synthase kinase-3 beta (GSK- catenin in the nucleus and stabilized E-cadherin to stabilize the intercellular junction and inhibit cell migration and growth. ^{24, 25)} also bridges in studies using medulloblastoma cells derived from a combination of Slit2 and Robo1 inhibited the cell migration ability by suppressing the actin cytoskeleton regulatory elements such as Cdc42 ^²⁶⁾ (Fig. 1) via the Rho GTPase. However, in one study using colon cancer-derived cell lines, increased expression of Slit2-Robo1 induced the activation of ubiquitin conjugated proteins, promoting the degradation of E-cadherin, and promoting cell growth and epithelial-mesenchmal transition The results were completely opposite. ^27). Therefore, the role of Slit and Robo is thought to be different depending on the types of cancer cells and their stages. ¹⁵⁾

현재 갑상선암에서 Slit-Robo의 발현 정도와 그 것이 갑상선암에 미치는 영향에 대해서는 거의 알려진 것이 없다. 하지만 최근 갑상선암의 빈도가 높은 가족을 대상으로 전장유전체 연관분석을 시행하여 연관성을 분석하였을 때 Slit-Robo Rho GTPase activating protein 1 (srGAP1) 유전자의 intron4에 위치한 SNP rs2168411이 갑상선 유두암의 감수성과 연관되어있음이 보고된바 있어 Slit-Robo 경로가 갑상선암의 발생에서도 중요한 역할이 있을 가능성을 시사하였다.²⁸⁾이에 본 연구는 갑상선암 세포주와 갑상선암 조직을 이용하여 Slit-Robo 경로가 갑상선암의 성장과 침습에 미치는 영향을 분석해보고자 하며, 이를 통해 새로운 바이오마커 및 새로운 치료 표적으로서 Slit-Robo 경로의 가능성을 확인해보고자 한다. Currently, little is known about the extent of Slit-Robo expression in thyroid cancer and its effect on thyroid cancer. However, when SNP rs2168411 located in the intron4 of the Slit-Robo Rho GTPase activating protein 1 (srGAP1) gene was associated with susceptibility to thyroid papillary carcinoma, , Suggesting that the Slit-Robo pathway may play an important role in the development of thyroid cancer. ^The aim of this study was to investigate the effect of the Slit-Robo pathway on the growth and invasion of thyroid cancer using thyroid carcinoma cell line and thyroid carcinoma tissue, and to confirm the possibility of the new biomarker and Slit-Robo pathway as a new therapeutic target I want to see.

본 명세서 전체에 걸쳐 다수의 논문 및 특허문헌이 참조되고 그 인용이 표시되어 있다. 인용된 논문 및 특허 문헌의 개시 내용은 그 전체로서 본 명세서에 참조로 삽입되어 본 발명이 속하는 기술 분야의 수준 및 본 발명의 내용이 보다 명확하게 설명된다.
Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

본 발명자들은 새로운 바이오마커 및 새로운 분자치료 표적을 이용한 갑상선암 전이 억제용 조성물을 개발하고자 예의 연구 노력하였다. 그 결과, 본 발명자들은 Slit-Robo 경로의 활성 저하가 종양 크기, 림프절 전이, 원격전이 및 재발 등의 갑상선암의 나쁜 예후와 연관 있음을 확인하였다. 또한, Slit의 투여로서 갑상선암 세포주의 성장 및 이동능이 뚜렷이 억제됨을 확인하였으며 이는 베타-카테닌(beta-catenin) 활성의 억제 및 E-카데린(E-cadherin) 활성 유도에 의한 것임을 규명하였다. 이로써 본 발명자들은 최초로 갑상선암에서 종양 억제 인자로서의 Slit-Robo 경로의 역할을 확인하였고, 새로운 바이오마커 및 새로운 분자 치료 표적으로서 Slit2 및 Robo1의 가능성을 확인함으로써 본 발명을 완성하게 되었다.The present inventors sought to develop a novel biomarker and a composition for inhibiting thyroid cancer metastasis using a novel molecular therapeutic target. As a result, the present inventors confirmed that the decrease in the activity of the Slit-Robo pathway is associated with a bad prognosis of thyroid cancer such as tumor size, lymph node metastasis, distant metastasis and relapse. In addition, the administration of Slit significantly inhibited the growth and migration of the thyroid carcinoma cell line, confirming the inhibition of beta-catenin activity and induction of E-cadherin activity. Thus, the present inventors first confirmed the role of the Slit-Robo pathway as a tumor suppressor in thyroid cancer, and confirmed the possibility of Slit2 and Robo1 as new biomarkers and novel molecular therapeutic targets, thus completing the present invention.

따라서, 본 발명의 목적은 원발성 갑상선암 전이 예방 또는 억제용 약제학적 조성물을 제공하는 데 있다.
Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or inhibiting primary thyroid cancer metastasis.

본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.
Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

본 발명의 일 양태에 따르면, 본 발명은 (a) 유효성분으로서 인간 SLIT2 (Slit homolog 2) 단백질 또는 유전자; 및 (b) 약제학적으로 허용되는 담체를 포함하는 원발성 갑상선암 전이 예방 또는 억제용 약제학적 조성물을 제공한다.
According to one aspect of the present invention, there is provided a human SLIT2 (Slit homolog 2) protein or gene as an active ingredient. And (b) a pharmaceutically acceptable carrier. The present invention also provides a pharmaceutical composition for preventing or inhibiting metastasis of primary thyroid cancer.

본 발명자들은 새로운 바이오마커 및 새로운 분자치료 표적을 이용한 갑상선암 전이 억제용 조성물을 개발하고자 예의 연구 노력한 결과, Slit-Robo 경로의 활성 저하가 종양 크기, 림프절 전이, 원격전이 및 재발 등의 갑상선암의 나쁜 예후와 연관 있음을 확인하였다. 또한, Slit의 투여로서 갑상선암 세포주의 성장 및 이동능이 뚜렷이 억제됨을 확인하였으며 이는 베타-카테닌(beta-catenin) 활성의 억제 및 E-카데린(E-cadherin) 활성 유도에 의한 것임을 규명하였다. 이로써 본 발명자들은 최초로 갑상선암에서 종양 억제 인자로서의 Slit-Robo 경로의 역할을 확인하였고, 새로운 바이오마커 및 새로운 분자 치료 표적으로서 Slit2 및 Robo1의 가능성을 확인하였다.The present inventors have made intensive studies to develop a composition for inhibiting thyroid cancer metastasis using a novel biomarker and a novel molecular therapy target. As a result, the inventors of the present invention found that the decrease in the activity of the Slit-Robo pathway leads to a bad prognosis of thyroid cancer including tumor size, lymph node metastasis, distant metastasis, . In addition, the administration of Slit significantly inhibited the growth and migration of the thyroid carcinoma cell line, confirming the inhibition of beta-catenin activity and induction of E-cadherin activity. The present inventors first confirmed the role of the Slit-Robo pathway as a tumor suppressor in thyroid cancer and confirmed the possibility of Slit2 and Robo1 as novel biomarkers and novel molecular therapeutic targets.

Slit-Robo 경로는 신경계 발생 과정에서 처음 발견되었으나 신경계 외에서도 다양한 세포의 성장과 이동능 등의 기능을 조절하는 것으로 생각되고 있다. 이에 Slit-Robo 경로가 정상 세포의 암성 변화를 유발하는 중요 원인일 가능성이 제기되었는데 실제로 신장암, 췌장암, 대장암 등에서 이들의 발현이 감소함이 보고된 바 있고 최근 연구는 췌장암에서 Slit-Robo경로의 발현 감소가 나쁜 예후와 연관이 있음을 발표하였다. 현재까지 갑상선암에서 Slit-Robo 경로의 발현 정도와 그 것이 갑상선암에 미치는 영향에 대해서는 거의 알려진 것이 없는데, 최근 갑상선 유두암을 대상으로 한 전장유전체 연관분석에서 Slit-Robo Rho GTPase activation protein 1 (SRGAP1) 유전자의 변이가 갑상선 유두암 감수성에 영향을 주는 원인으로 밝혀져 갑상선암에서도 Slit-Robo 경로가 중요한 역할을 할 것을 시사하였다. 이에 본 연구에서는 갑상선암 세포주와 갑상선암 조직을 이용하여 Slit-Robo 경로가 갑상선암의 성장과 침습에 미치는 영향을 다각적으로 분석해 보고자 한다.The Slit-Robo pathway was first found in the development of the nervous system, but it is thought to regulate various cell growth and migration functions outside the nervous system. In this study, we investigated the role of Slit-Robo pathway in the pathogenesis of pancreatic carcinoma. We investigated the role of Slit-Robo pathway in the pathogenesis of pancreatic cancer. Of the patients had a poor prognosis. The role of the Slit-Robo Rho GTPase activation protein 1 (SRGAP1) gene in thyroid papillary carcinoma is now well documented. The mutation was found to be a causative factor for papillary thyroid cancer susceptibility, suggesting that Slit-Robo pathway plays an important role in thyroid cancer. The purpose of this study was to analyze the effect of Slit-Robo pathway on growth and invasion of thyroid cancer using thyroid cancer cell line and thyroid cancer tissue.

원발성 갑상선암(primary thyroid cancer)의 대부분은 분화암으로서 대표적으로 갑상선 유두암과 갑상선 여포암이 있다. 이 중 갑상선 유두암은 전체 갑상선암의 60%-70%를 차지하는 암으로서, 어느 연령에나 발생하지만 20대-30대에서 주로 발생한다. 갑상선 유두암은 주로 림프계로 전이 되지만 전체적으로 매우 천천히 자라고 10년 생존율이 90%이상으로 예후가 좋은 암이지만, 드물게 폐나 뼈 등으로 원격전이하는 예가 있어 초기치료가 중요하게 여겨진다. Primary thyroid cancer is mostly differentiated from thyroid papillary cancer and thyroid follicular cancer. Thyroid papillary cancer is a cancer that accounts for 60% -70% of all thyroid cancer, occurring in any age but mainly in the 20s-30s. Thyroid papillary carcinoma is mainly metastatic to the lymphatic system. However, the overall prognosis is good, with a 10-year survival rate of more than 90%. However, early metastasis is very important because of distant metastasis to the lung or bone.

본 발명의 일 예에 따르면, 상기 갑상선암은 갑상선 유두암이다.According to one embodiment of the present invention, the thyroid cancer is thyroid papillary cancer.

본 발명의 검출대상인 인간 SLIT2 (Slit homolog 2) 단백질은 서열목록 제1서열(NP_004778.1; 1529 aa), 서열목록 제2서열(NP_001276064; 1525 aa) 또는 서열목록 제3서열(NP_001276065.1; 1521aa) 의 아미노산 서열을 포함하며, 상기 아미노산 서열은 각각 서열목록 제4서열(NM_004787.2; 6398 bp), 서열목록 제5서열(NM_001289135.1; 6386 bp) 또는 서열목록 제6서열(NM_001289136.1; 6374 bp) 의 뉴클레오타이드 서열에 의해 코딩된다. 한편, 인간 ROBO1 (Roundabout homolog 1) 단백질은 서열목록 제7서열(NP_002932.1; 1651 aa), 서열목록 제8서열(NP_598334.2; 1606 aa) 또는 서열목록 제9서열(NP_001139317.1; 1551 aa) 의 아미노산 서열을 포함하며, 상기 아미노산 서열은 각각 서열목록 제10서열(NM_002941.3; 6895 bp), 서열목록 제11서열(NM_133631.3; 7550 bp) 및 서열목록 제12서열(NM_001145845.1 (7385 bp) 의 뉴클레오타이드 서열에 의해 코딩된다. The human SLIT2 (Slit homolog 2) protein to be detected in the present invention may be selected from the group consisting of SEQ ID NO: 1 (NP_004778.1; 1529 aa), SEQ ID NO: 2 (NP_001276064; 1525 aa) or SEQ ID NO: 3 (NP_001276065.1; (NM_004787.2; 6398 bp), SEQ ID NO: 5 (NM_001289135.1; 6386 bp), or the amino acid sequence of SEQ ID NO: 6 (NM_001289136. 1; 6374 bp). On the other hand, the human ROBO1 (Roundabout homolog 1) protein may be selected from the group consisting of SEQ ID NO: 7 (NP_002932.1; 1651 aa), SEQ ID NO: 8 (NP_598334.2; 1606 aa) or SEQ ID NO: 9 (NP_001139317.1; wherein said amino acid sequence is selected from the group consisting of SEQ ID NO: 10 (NM_002941.3; 6895 bp), SEQ ID NO: 11 (NM_133631.3; 7550 bp) and SEQ ID NO: 12 (NM_001145845. 1 (7385 bp) nucleotide sequence.

본 발명의 일 예에 따르면, 본 발명의 약제학적 조성물은 (a) 서열목록 제1서열, 서열목록 제2서열 또는 서열목록 제3서열의 아미노산 서열을 포함하는 인간 SLIT2 (Slit homolog 2) 단백질, 또는 상기 아미노산 서열을 코딩하는 뉴클레오타이드 서열을 유효성분으로 포함한다.According to one embodiment of the present invention, the pharmaceutical composition of the present invention comprises (a) a human SLIT2 (Slit homolog 2) protein comprising the amino acid sequence of SEQ ID No. 1, SEQ ID No. 2 or SEQ ID No. 3, Or a nucleotide sequence encoding said amino acid sequence as an active ingredient.

보다 상세하게는, 본 발명의 약제학적 조성물은 (a) 서열목록 제1서열, 서열목록 제2서열 또는 서열목록 제3서열의 아미노산 서열을 포함하는 인간 SLIT2 (Slit homolog 2) 단백질 또는 서열목록 제1서열, 서열목록 제2서열 또는 서열목록 제3서열의 아미노산 서열을 코딩하는 뉴클레오타이드 서열을 포함하는 유전자 전달체를 유효성분으로 포함한다. More particularly, the pharmaceutical composition of the present invention comprises (a) a human SLIT2 (Slit homolog 2) protein comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3, 1 sequence, a second sequence, or a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 3 as an active ingredient.

본 발명에서 “SLIT2 유전자”는 SLIT2 단백질을 코딩하는 전체 뉴클레오타이드 서열 뿐만 아니라, 갑상선암 전이 억제 활성을 나타내는 SLIT2 단백질의 단편 또는 서브유닛(subunit)을 코딩하는 뉴클레오타이드 서열도 포함하는 의미로 해석한다. In the present invention, " SLIT2 gene " means not only the entire nucleotide sequence encoding the SLIT2 protein, but also a nucleotide sequence encoding a fragment or subunit of the SLIT2 protein exhibiting thyroid cancer metastasis inhibitory activity.

본 발명에서 상기 핵산 서열을 운반하는 상기 유전자 전달체(gene carrier)는 당업계에 공지된 다양한 유전자 전달체를 이용할 수 있으며, 예컨대, 플라스미드, 파아지 등을 포함하며, 바람직하게는 상기 유전자 전달체는 플라스미드이다.In the present invention, the gene carrier carrying the nucleic acid sequence may use various gene carriers known in the art, for example, a plasmid, a phage, etc. Preferably, the gene carrier is a plasmid.

본 발명에서 이용되는 유전자 전달체는 인간 SLIT2 단백질 을 코딩하는 서열목록 제4서열, 서열목록 제5서열 또는 서열목록 제6서열의 뉴클레오타이드 서열을 포함하는 발현 컨스트럭트를 포함한다. The gene carrier used in the present invention includes an expression construct comprising a nucleotide sequence of SEQ ID No. 4, SEQ ID No. 5, or SEQ ID No. 6 of the human SLIT2 protein.

본 발명의 발현 컨스트럭트는 당업계에 공지된 다양한 방법을 통해 구축될 수 있으며, 이에 대한 구체적인 방법은 Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press(2001)에 개시되어 있으며, 이 문헌은 본 명세서에 참조로서 삽입된다.The expression constructs of the present invention can be constructed through various methods known in the art, and specific methods for this are disclosed in Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press (2001) , Which is incorporated herein by reference.

본 명세서에서 용어 “뉴클레오타이드”는 DNA 그리고 RNA 분자를 포괄적으로 포함하는 의미를 갖으며, 자연의 뉴클레오타이드뿐만 아니라, 당 또는 염기 부위가 변형된 유사체(analogue)도 포함한다(Scheit, Nucleotide Analogs, John Wiley, New York(1980); Uhlman 및 Peyman, Chemical Reviews, 90:543-584(1990)).As used herein, the term " nucleotide " is meant to encompass DNA and RNA molecules, including natural nucleotides as well as analogues in which sugar or base regions are modified (Scheit, Nucleotide Analogs, John Wiley , New York (1980); Uhlman and Peyman, Chemical Reviews, 90: 543-584 (1990)).

본 발명의 유전자 전달체에서 상기 SLIT2 단백질을 코딩하는 뉴클레오타이드 서열은 상기 서열목록 제1서열, 서열목록 제2서열 또는 서열목록 제3서열로 표시되는 뉴클레오타이드 서열에 대하여 실질적인 동일성을 나타내는 뉴클레오타이드 서열도 포함한다. 실질적인 동일성은, 상기한 본 발명의 뉴클레오타이드 서열과 임의의 다른 서열을 최대한 대응되도록 얼라인하고, 당업계에서 통상적으로 이용되는 알고리즘을 이용하여 얼라인된 서열을 분석한 경우에, 최소 80%의 상동성, 보다 바람직하게는 최소 90%의 상동성, 가장 바람직하게는 최소 95%의 상동성을 나타내는 뉴클레오타이드 서열을 의미한다.The nucleotide sequence coding for the SLIT2 protein in the gene carrier of the present invention also includes a nucleotide sequence which shows substantial identity to the nucleotide sequence shown in the above Sequence Listing first sequence, Sequence Listing second sequence or Sequence Listing third sequence. Substantial identity is determined by aligning the nucleotide sequence of the present invention with any other sequence as closely as possible and analyzing the aligned sequence using algorithms commonly used in the art to identify at least 80% Homology, more preferably at least 90% homology, and most preferably at least 95% homology.

한편, 상기 발현 컨스트럭트가 플라스미드 벡터인 경우, 상기 SLIT2 단백질 코딩 뉴클레오타이드 서열은 프로모터에 작동적으로 연결되는 것이 바람직하다. 이 경우의 발현 컨스트럭트는 발현 목적의 폴리뉴클레오타이드 서열 및 이 서열의 발현을 유도하는 발현서열(예컨대, 프로모터)을 포함하는 발현을 위한 최소한의 엘리먼트(elements)를 포함한 구조를 의미한다. 이러한 발현 컨스트럭트는 전사조절 서열-발현 목적의 뉴클레오타이드 서열-폴리아데닐화 서열을 포함할 수 있다.On the other hand, when the expression construct is a plasmid vector, the SLIT2 protein coding nucleotide sequence is preferably operatively linked to a promoter. The expression construct in this case means a structure including a minimum element for expression including a polynucleotide sequence for expression purpose and an expression sequence (for example, a promoter) for inducing expression of this sequence. Such expression constructs may include a nucleotide sequence-polyadenylation sequence for transcriptional control sequence-expression purposes.

본 발명의 일 에 따르면, 본 발명의 상기 유전자 전달체는 (ⅰ) SLIT2 단백질을 코딩하는 핵산 분자 (ⅱ) 상기 핵산 분자에 작동적으로 결합(operatively linked)되어 있고 동물세포에서 RNA 분자를 형성시키는 프로모터; 및 (ⅲ) 동물세포에서 작용하여 RNA 분자의 3'-말단의 폴리아데닐화를 야기시키는 폴리 A 시그널 서열을 포함하는 동물세포 발현용 재조합 벡터이다.According to one aspect of the present invention, the gene delivery system of the present invention comprises (i) a nucleic acid molecule encoding a SLIT2 protein, (ii) a promoter operably linked to the nucleic acid molecule, ; And (iii) a poly A signal sequence that acts in animal cells to cause polyadenylation of the 3'-end of the RNA molecule.

본 명세서에서 용어 “프로모터”는 코딩 서열 또는 기능적 RNA의 발현을 조절하는 DNA 서열을 의미한다. 본 발명의 발현 벡터는 재조합 발현 벡터로서 발현대상물질-코딩 뉴클레오타이드 서열은 상기 프로모터에 작동적으로 연결된다. 본 명세서에서 용어 “작동적으로 결합된(operatively linked)”은 핵산 발현 조절 서열(예: 프로모터 서열, 시그널 서열, 또는 전사조절인자 결합 위치의 어레이)과 다른 핵산 서열 사이의 기능적인 결합을 의미하며, 이에 의해 상기 조절 서열은 상기 다른 핵산 서열의 전사 및/또는 번역을 조절하게 된다. As used herein, the term " promoter " means a DNA sequence that regulates the expression of a coding sequence or functional RNA. The expression vector of the present invention is a recombinant expression vector, and the substance-coding nucleotide sequence to be expressed is operatively linked to the promoter. As used herein, the term " operatively linked " refers to a functional linkage between a nucleic acid expression control sequence (e.g., an array of promoter sequences, signal sequences, or transcription factor binding sites) , Whereby the regulatory sequence regulates transcription and / or translation of the other nucleic acid sequence.

본 발명에 있어서 프로모터는 동물세포에서 발현하기에 적합한 프로모터가 선택된다. 예컨대, CMV(Cytomegalovirus) 프로모터, 아데노바이러스 후기 프로모터, 백시니아 바이러스 7.5K 프로모터, SV40(Simian Virus 40) 프로모터, SV40E1 프로모터, HSV(Herpes simplex virus)의 tk 프로모터, RSV(Respiratory syncytial virus) 프로모터, EF1(elongation factor-1 alpha) 알파 프로모터, 메탈로티오닌 프로모터, 베타-액틴 프로모터, 인간 IL-2(interluekin-2) 유전자의 프로모터, 인간 IFN(interferon) 유전자의 프로모터, 인간 IL-4 유전자의 프로모터, 인간 림포톡신 유전자의 프로모터 또는 인간 GM-CSF(Granulocyte macrophage colony-stimulating factor) 유전자의 프로모터가 사용될 수 있다.As the promoter in the present invention, a promoter suitable for expression in animal cells is selected. Examples of such promoters include CMV (cytomegalovirus) promoter, adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 (Simian Virus 40) promoter, SV40E1 promoter, HSV (herpes simplex virus) tk promoter, RSV (respiratory syncytial virus) promoter, EF1 promoter, beta-actin promoter, human IL-2 (interluekin-2) gene promoter, human IFN (interferon) gene promoter, human IL-4 gene promoter , A promoter of human lymphotoxin gene or a promoter of human GM-CSF (Granulocyte macrophage colony-stimulating factor) gene can be used.

본 발명에 있어서 벡터는 전사 종결 서열로서 폴리아데닐화 서열을 포함하며, 소성장 호르몬 터미네이터, HSV(Herpes simplex virus) 유래 TK(Thymidine kinase) 또는 SV40(Simian virus 40) 유래 폴리 아데닐화 서열이 바람직하다.In the present invention, the vector includes a polyadenylation sequence as a transcription termination sequence, and is preferably a bovine growth hormone terminator, a herpes simplex virus derived TK (Thymidine kinase) or a SV40 (Simian virus 40) derived polyadenylation sequence .

본 발명에서 이용되는 벡터는 선택마커로서 당업계에서 통상적으로 이용되는 동물세포에 작용하는 항생제 내성 유전자를 포함할 수 있으며, 예를 들어 제네티신(G418), 퓨로마이신, 암피실린, 겐타마이신, 카베니실린, 클로람페니콜, 스트렙토마이신, 카나마이신, 네오마이신 및 테트라사이클린에 대한 내성 유전자가 있다. The vector used in the present invention may include an antibiotic resistance gene which acts on an animal cell which is commonly used in the art as a selection marker and includes, for example, geneticin (G418), puromycin, ampicillin, gentamicin, There is a resistance gene for benicillin, chloramphenicol, streptomycin, kanamycin, neomycin and tetracycline.

갑상선암 전이는 크게 4 단계로 나뉠 수 있으며, 암 조직이 주변 림프절 전이 없이 갑상선 조직 일부를 침범한 단계(제1기); 암조직이 주변 림프절로 전이하지는 않았으나 갑상선 조직 전체를 침범한 단계(제2기); 주변 림프절로의 전이가 있는 단계(제3기); 및 다른 신체부위로 전이되는 원격전이(distant metastasis)가 있는 단계(제4기)로 나뉠 수 있다. 본 발명의 조성물은 제3기 및 제4기의 갑상선암에서 림프절 전이 및 원격전이를 효과적으로 억제할 수 있다. Thyroid carcinoma metastasis can be divided into four stages: the stage where cancer tissue invades part of the thyroid tissue without surrounding lymph node metastasis (stage 1); Stage in which the cancerous tissue did not migrate to the surrounding lymph nodes but invaded the entire thyroid tissue (stage 2); Stage with metastasis to surrounding lymph nodes (stage 3); And a stage with distant metastasis that is metastasized to other parts of the body (stage 4). The composition of the present invention can effectively inhibit lymph node metastasis and distant metastasis in the third and fourth stage of thyroid cancer.

본 발명의 조성물은 갑상선 종양의 크기, 림프절 전이 발생율, 원격전이 발생율 또는 재발율을 감소시킨다. 예컨대, 림프절 전이 또는 원격전이가 나타난 원발성 갑상선암 환자에 본 발명의 조성물을 투여할 경우, 갑상선 종양의 크기가 감소하고, 림프절 전이 및 원격전이 현상을 차단 또는 감소시킬 수 있으며, 갑상선암의 재발 발생율을 낮추는 효과가 있다.The compositions of the present invention reduce the size of the thyroid tumor, the incidence of lymph node metastasis, the rate of metastasis or the recurrence rate. For example, administration of a composition of the present invention to a primary thyroid cancer patient who has lymph node metastasis or distant metastasis may reduce the size of the thyroid tumor, block or reduce the lymph node metastasis and distant metastasis, and decrease the incidence of recurrence of thyroid cancer It is effective.

또한, 본 발명의 갑상선암 전이 억제 조성물은 Wnt 신호경로의 하위 조절자의 발현을 감소시키고, 특히 베타카테닌의 발현을 억제한다. 이로써 E-카데린(E-cadherin)의 발현을 증가시켜 암 세포의 전이를 억제한다. In addition, the thyroid cancer metastasis inhibiting composition of the present invention reduces the expression of sub-regulators of the Wnt signal pathway, and particularly inhibits the expression of beta-catenin. This increases the expression of E-cadherin and inhibits metastasis of cancer cells.

본 발명의 구체적인 일 실시예에 따르면, 본 발명자들은 갑상선 조직미세배열 슬라이드에서 Slit2와 Robo1의 면역화학염색을 시행하였을 때 Slit2 및 Robo1 모두 정상 또는 양성 조직 보다 갑상선 유두암에서 발현이 뚜렷하게 증가하는 것을 관찰하였는데(도 6 및 도 7 참조), 림프절 전이 또는 원격전이가 있는 경우에는 Slit2와 Robo1의 발현이 오히려 감소함을 확인하였다. 이러한 차이는 Slit2 에서 더 뚜렷하게 관찰되었다. Slit-Robo 발현과 환자의 임상경과의 연관성을 분석하였을 때 Slit2의 발현 저하는 림프절 전이 및 원격전이, 재발 위험 증가와 유의한 연관을 보였고, 이러한 연관성은 BRAF 변이 유무와는 무관하였다(표 1 및 표 2 참조). 또한 Slit2와 Robo1의 발현이 모두 양성인 환자와 비교하여 그렇지 않은 환자들의 종양 크기가 유의하게 큰 경향도 관찰할 수 있었다. According to one embodiment of the present invention, the immunohistochemical staining of Slit2 and Robo1 in a thyroid microarray slide revealed that Slit2 and Robo1 showed markedly increased expression in papillary thyroid cancer than in normal or benign tissues (See FIGS. 6 and 7), and that the expression of Slit2 and Robo1 was decreased in the presence of lymph node metastasis or distant metastasis. This difference was more pronounced in Slit2. When Slit-Robo expression was correlated with the patient's clinical course, the decreased expression of Slit2 was significantly associated with increased risk of lymph node metastasis, distant metastasis, and recurrence, and this association was independent of BRAF mutations (Table 1 and Table 2) See Table 2). In addition, the tumor size was significantly larger in patients with both Slit2 and Robo1 than in those with positive expression of Slit2 and Robo1.

한편, Slit2 와 Slit3 를 갑상선암 세포주에 처리하였을 때 농도 의존적인 세포 성장 억제가 관찰되었고 특히, 갑상선 유두암 유래 세포주인 BCPAP와 TPC-1에서 뚜렷한 차이를 보였다. 이 중 BCPAP 세포주를 이용하여 상처치유분석 및 transwell chamber 침습검사를 시행한 결과, Slit의 세포 이동능 억제 효과를 확인할 수 있었으며, Slit 투여 후 beta-catenin의 활성이 억제되고, E-cadherin 활성이 유도되며 wnt3A 투여로 활성화되는 beta-catenin 전사인자 활성이 억제됨을 확인하였다. On the other hand, when Slit2 and Slit3 were treated with thyroid cancer cell line, a concentration-dependent inhibition of cell growth was observed. In particular, there was a clear difference between BCPAP and TPC-1, the papillary thyroid cancer cell lines. In the BCPAP cell line, wound healing analysis and transwell chamber invasion test showed that Slit inhibited the cell migration, inhibited beta-catenin activity, and E-cadherin activity And the activity of beta-catenin transcription factor activated by wnt3A was inhibited.

본 발명의 약제학적 조성물은 약제학적으로 허용되는 담체를 포함한다. 본 발명의 약제학적 조성물에 포함되는 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약제학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. 적합한 약제학적으로 허용되는 담체 및 제제는 Remington's Pharmaceutical Sciences (19th ed., 1995)에 상세히 기재되어 있다.The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 약제학적 조성물의 1일 투여량은 예컨대 0.001-1000 ㎎/㎏이다. 그러나, 유효 성분의 실제 투여량은 분화 및 증식하고자 하는 대상조직 세포의 양, 투여경로, 환자의 체중, 연령 및 성별 등 여러 관련 인자를 고려하여 결정할 수 있으며, 따라서, 상기 투여량은 어떠한 형태로든 본 발명의 범위를 한정하는 것은 아니다.
The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . The daily dosage of the pharmaceutical composition of the present invention is, for example, 0.001-1000 mg / kg. However, the actual dose of the active ingredient may be determined taking into account various relevant factors such as the amount of target tissue cells to be differentiated and proliferated, the route of administration, the body weight of the patient, age and sex, The scope of the present invention is not limited thereto.

본 발명의 특징 및 이점을 요약하면 다음과 같다: The features and advantages of the present invention are summarized as follows:

(a) 본 발명은 원발성 갑상선암 전이 예방 또는 억제용 약제학적 조성물에 관한 것이다.(a) The present invention relates to a pharmaceutical composition for the prevention or inhibition of primary thyroid cancer metastasis.

(b) 본 발명자들은 SLIT-Robo 경로의 활성 저하는 큰 종양 크기, 림프절 전이, 원격전이와 재발 등의 갑상선암의 나쁜 예후와 연관 있음을 최초로 규명하였다.(b) The inventors of the present invention for the first time have found that the deactivation of the SLIT-Robo pathway is associated with a bad prognosis of thyroid cancer including large tumor size, lymph node metastasis, distant metastasis and recurrence.

(c) 본 발명자들은 SLIT2-Robo1 발현율 측정을 이용하여 원발성 갑상선암 전이여부를 진단할 수 있는 방법을 제안한다.(c) The present inventors propose a method for diagnosing primary thyroid cancer metastasis using the SLIT2-Robo1 expression rate measurement.

(d) 또한, 본 발명은 최초로 갑상선암에서 종양 억제 인자로서의 SLIT-Robo 경로의 역할을 확인하였으며, 새로운 바이오마커 및 새로운 분자 치료 표적으로 SLIT2의 가능성을 제시한다.
(d) In addition, the present invention first confirmed the role of the SLIT-Robo pathway as a tumor suppressor in thyroid cancer, suggesting the possibility of SLIT2 as a new biomarker and novel molecular therapy target.

도 1은 SLIT2-Robo1 경로 활성에 따른 세포 내 신호체계 변화 모식도이다.
*β-cat, beta-catenin; PI3K, phosphatidyl inositol 3-kinase; Akt, v-akt murine thymoma viral oncogene; GSK3 β, glycogen synthase kinase-3 beta; srGAP, SLIT-Robo Rho GTPase activating protein; E-cad, E-cadherin.
도 2는 갑상선 조직 미세 슬라이드에서 SLIT2 면역화학염색 결과의 예를 나타낸 것이다. A/C/E/F는 SLIT2 발현 음성의 예이고, B/D/G/H는 SLIT2 발현 양성의 예시이다. Normal : 대응되는 정상 조직; NH :결절성과증식증; FA :여포성 선종; PTC :갑상선 유두암.
도 3은 갑상선 조직 미세 슬라이드에서 Robo1 면역화학염색 결과의 예를 나타낸 것이다. A/B/D/E/F는 Robo1 발현 음성의 예이고 C/G/H는 Robo1 발현 양성의 예시이다. Normal : 대응되는 정상 조직; NH :결절성과증식증; FA :여포성 선종; PTC :갑상선 유두암
도 4a 및 도 4b는 갑상선암 세포주의 SLIT-Robo 발현정도를 나타낸 것이다. 모든 Robo, SLIT 아형의 발현을 18S 발현과 비교하여 ΔCT value를 구하였으며, 그 중 가장 ΔCT value가 큰 Robo 및 SLIT의 아형을 기준으로 하여 상대발현도(2^{-ΔΔCT value})을 나타내었다. (4a) Robo, (4b) SLIT.
도 5는 정상 및 갑상선암 신선동결조직에서 SLIT과 Robo 발현을 비교한 결과이다. 모든 Robo 및 SLIT 아형의 발현을 18S 발현과 비교하여 ΔCT value를 구하였으며, 정상 조직 중 가장 ΔCT value가 큰 Robo 및 SLIT의 아형을 기준으로 하여 상대발현도(2^{-ΔΔCT value})을 나타내었다.
도 6은 갑상선 조직형에 따른 Robo1 면역화학염색 양성률을 비교한 결과이다. 대응되는 정상 조직과 양성 갑상선 종양에 비해 갑상선 유두암에서 현저한 Robo1 양성률 증가를 나타내었다. Benign: 양성 갑상선 종양 ; PTC : 갑상선 유두암; Normal : 대응되는 정상 조직.
도 7은 갑상선 조직형에 따른 SLIT2 면역화학염색 양성률을 비교한 결과이다. 대응되는 정상 조직과 양성 갑상선 종양에 비해 갑상선 유두암에서 현저한 SLIT2 양성률 증가를 나타내었다. Benign: 양성 갑상선 종양 ; PTC : 갑상선 유두암; Normal : 대응되는 정상 조직.
도 8은 Robo1 및 SLIT2 면역화학염색 양성률과 림프절 전이 및 원격 전이의 관련성을 비교한 것이다. 갑상선 유두암에서 림프절 전이가 동반되면 Robo1 및 SLIT2 양성률이 의미 있게 감소하였다. 또한 원격전이의 동반은 SLIT2 양성률 감소와 뚜렷한 연관성을 나타내었다.
도 9는 Robo1/SLIT2 발현에 따른 무재발 생존율을 비교한 결과이다. SLIT2 발현 감소는 현저한 무재발 생존 악화와 통계적으로 유의한 연관성을 나타내었다.
도 10은 SLIT 투여에 따른 갑상선암 세포주의 성장 변화를 나타낸 결과이다(세포 수 측정). BCPAP 와 TPC-1 세포주에서 농도 의존적인 성장 감소 효과가 관찰되었다.
도 11은 SLIT 투여에 따른 갑상선암 세포주의 성장 변화를 나타낸 결과이다(BrdU 측정). BCPAP와 TPC-1 세포주에서 농도 의존적인 성장 감소 효과가 관찰되었으며 세포 수 측정 결과와 유사한 결과를 나타내었다.
도 12는 SLIT 투여 후 갑상선암 세포주의 이동능 변화를 나타낸 결과이다(상처치유분석). BCPAP 세포주에 SLIT2와 SLIT3 3nM을 투여하고 24시간 관찰하였을 때 대조군에 비해 현저한 상처치유억제가 관찰되었다.
도 13은 SLIT 투여 후 갑상선암 세포주의 이동능 변화를 나타낸 결과이다(Transwell chamber 침습분석). 대조군과 비교하여, BCPAP 세포주에 SLIT2와 SLIT3를 투여하였을 때 아래 공간으로 침습한 세포의 수 및 흡광도가 모두 현저히 감소하였다.
도 14는 SLIT 투여 후 분자신호전달체계의 변화를 나타낸다. SLIT2 및 SLIT3 투여시 활성(active) 베타카테닌의 발현이 감소하고 E-cadherin의 발현이 증가하는 것을 확인하였다.
도 15는 SLIT 투여 후 Wnt/beta-catenin 활성도의 변화를 나타낸다. SLIT 투여는 STF(Super 19X TOPFlash) 형질전환 세포에서 Wnt3A 투여로 활성된 LEF/TCF 전사를 의미 있게 억제하였다(*p < 0.01).FIG. 1 is a schematic diagram of an intracellular signaling system according to SLIT2-Robo1 pathway activity. FIG.
* β-cat, beta-catenine; PI3K, phosphatidyl inositol 3-kinase; Akt, v-akt murine thymoma viral oncogene; GSK3?, Glycogen synthase kinase-3 beta; srGAP, SLIT-Robo Rho GTPase activating protein; E-cad, E-cadherin.
Figure 2 shows an example of the results of SLIT2 immunochemical staining on thyroid tissue microslides. A / C / E / F is an example of SLIT2 expression negative, and B / D / G / H is an example of SLIT2 expression positive. Normal: Corresponding normal tissue; NH: Nodular hyperplasia; FA; PTC: Thyroid papillary cancer.
Figure 3 shows an example of Robo1 immunochemical staining results on thyroid tissue microarray. A / B / D / E / F is an example of Robo1 expression negative and C / G / H is an example of positive Robo1 expression. Normal: Corresponding normal tissue; NH: Nodular hyperplasia; FA; PTC: Thyroid papilloma
4A and 4B show SLIT-Robo expression levels of thyroid cancer cell lines. The expression of all Robo and SLIT subtypes was compared with that of 18S expression, and the relative expression level (2 ^{-ΔΔCT value} ) was ^calculated based on subtypes of Robo and SLIT with the ^{highest ΔCT value} . (4a) Robo, (4b) SLIT.
FIG. 5 shows the comparison of SLIT and Robo expression in fresh frozen tissues of normal and thyroid cancer. Expression of all Robo and SLIT subtypes was compared with 18S expression, and ^{ΔCT value} was calculated. Relative expression (2- ^{ΔΔCT value} ) was shown based on subtypes of Robo and SLIT with the highest ΔCT value among normal tissues.
FIG. 6 shows the results of comparing Robo1 immunochemistry staining according to thyroid tissue type. There was a significant increase in the positive rate of Robo1 in papillary thyroid carcinoma compared with normal tissue and benign thyroid tumor. Benign: benign thyroid tumor; PTC: Thyroid papillary cancer; Normal: Corresponding normal tissue.
FIG. 7 shows the results of comparing the positive rate of SLIT2 immunochemical staining according to thyroid tissue type. The positive rate of SLIT2 was increased in papillary thyroid carcinoma compared with normal tissue and benign thyroid tumor. Benign: benign thyroid tumor; PTC: Thyroid papillary cancer; Normal: Corresponding normal tissue.
Figure 8 compares the relationship between Robo1 and SLIT2 immunochemistry stain positive rate and lymph node metastasis and distant metastasis. When lymph node metastasis was present in papillary thyroid carcinoma, the positive rate of Robo1 and SLIT2 decreased significantly. The association of distant metastasis was also significantly associated with a decrease in SLIT2 positivity.
FIG. 9 shows the results of comparing the recurrence-free survival rate according to Robo1 / SLIT2 expression. The decrease in SLIT2 expression was statistically significant in relation to the significant recurrence-free survival.
FIG. 10 shows the growth of thyroid cancer cell line according to SLIT administration (cell count). BCPAP and TPC-1 cell lines showed a concentration-dependent growth-reducing effect.
Fig. 11 shows the growth of thyroid cancer cell line according to administration of SLIT (BrdU measurement). In the BCPAP and TPC-1 cell lines, a concentration-dependent growth-reducing effect was observed and the results were similar to those of the cell counts.
FIG. 12 shows the change in the migration ability of the thyroid cancer cell line after SLIT administration (wound healing analysis). When SLIT2 and 3 nM of SLIT3 were administered to BCPAP cell line for 24 hours, remarkable wound healing inhibition was observed compared to the control group.
FIG. 13 shows the change in the migration of thyroid cancer cell line after administration of SLIT (Transwell chamber invasion assay). When SLIT2 and SLIT3 were administered to the BCPAP cell line, the numbers of the cells invading into the lower space and the absorbance were significantly decreased compared to the control group.
Figure 14 shows the change in the molecular signaling system after administration of SLIT. SLIT2 and SLIT3 decreased the expression of active beta-catenin and increased the expression of E-cadherin.
Figure 15 shows the change in Wnt / beta-catenin activity after administration of SLIT. SLIT administration significantly inhibited Wnt3A-induced LEF / TCF transcription in STF (Super 19X TOPFlash) transfected cells (* p <0.01).

이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

실시예Example

연구재료Research material

세포 배양에 필요한 열불활성화된 우태아 혈청, 페니실린-스트렙토마이신계 항생제, 인산염 완충용액 (PBS), 2.5% Trypsin-EDTA, RPMI-1640, DMEM 배지는 Gibco (Grand Island, NY, USA) 에서 구입하였다. SLIT2 단백과 조직의 면역화학염색에 이용된 Robo1 일차 항체, GAPDH 일차항체는 Abcam (Cambridge, MA, USA)에서 구입하였고 SLIT3 단백은 Abnova (Jhongli, Taiwan)에서 구입하였다. RNeasy Mini 키트와 QuantiFastSYBRGreen PCR 키트, QIAampDNA FFPE 조직 키트는 Qiagen (Valencia, CA, USA) 에서 구입하였다. 한천은 Bioline (Taunton, MA, USA)에서, 니트로셀룰로스막은 Amersham Bioscience (Piscataway, NJ, USA)에서 구입하였다. TRIzol RNA 분리시약, NuPAGE 겔, HRP 화학발광면역기질시약, LipofectamineTM 2000은 Invitrogen (Carlsbad, CA, USA)에서, RevertAid 첫가닥 상보적DNA 합성키트는 Fermentas (Glen Brunie, MD, USA)에서 구입하였다. Cell proliferation ELISA, BrdU kit는 Roche (Mannheim, Germany) 에서 구입하였다. 세포 침습능 분석을 위한 transwell chamber 키트는 Cell biolabs (San Diego, CA, USA) 에서 구입하였다. 단백질 정량에 사용된 Quanti-iTTM 단백 분석키트는 Molecular Prob (Eugene, OR, USA) 에서 구입하였다. 조직의 면역화학염색에 이용된SLIT2 일차 항체와 beta-catenin에 대한 일차항체는 Santa Cruz Biotechnology (Santa Cruz, CA, USA)에서 구입하였다. 활성화 beta-catenin에 대한 일차 항체는 EMD Milipore (Billerica, MA, USA)에서, E-cadherin에 대한 일차항체는 Cell signaling (Danver, MA, USA)에서 구입하였다. 이차항체는 Vector Laboratories (Burlingame, CA, USA)에서 구입하였다. 발광리포터 분석을 위한 이중 발광효소 키트는 Promega (Madison, WI, USA) 에서 구입하였다. 발광 리포터 분석을 위해 사용된 super 19X TOPFlash(STF)와 8XFOPFlash(SFF) 벡터 및 Renilla 발광효소 벡터, Wnt3A 분비 L 292세포는 Amiee Kohn의 실험실에서 구하였다.
2.5% trypsin-EDTA, RPMI-1640 and DMEM medium were purchased from Gibco (Grand Island, NY, USA) for the cell culture, and the fetal bovine serum, penicillin-streptomycin antibiotics, phosphate buffer solution . Robo1 primary antibody and GAPDH primary antibody used for immunohistochemical staining of SLIT2 protein and tissue were purchased from Abcam (Cambridge, MA, USA) and SLIT3 protein was purchased from Abnova (Jhongli, Taiwan). RNeasy Mini kit, QuantiFastSYBRGreen PCR kit, QIAampDNA FFPE tissue kit were purchased from Qiagen (Valencia, CA, USA). Agar was purchased from Bioline (Taunton, MA, USA) and nitrocellulose membranes from Amersham Bioscience (Piscataway, NJ, USA). RevertAid first strand complementary DNA synthesis kit was purchased from Fermentas (Glen Brunie, MD, USA) in Invitrogen (Carlsbad, CA, USA) and TRIzol RNA isolation reagent, NuPAGE gel, HRP chemiluminescent immune substrate reagent, Lipofectamine Cell proliferation ELISA and BrdU kit were purchased from Roche (Mannheim, Germany). The transwell chamber kit for cell invasion was purchased from Cell biolabs (San Diego, CA, USA). Quanti-iT ™ protein assay kit used for protein determination was purchased from Molecular Prob (Eugene, OR, USA). Primary antibodies against SLIT2 primary antibody and beta-catenin used for tissue immunochemical staining were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The primary antibody against activated beta-catenin was purchased from EMD Milipore (Billerica, MA, USA) and the primary antibody against E-cadherin was purchased from Cell signaling (Danver, MA, USA). Secondary antibodies were purchased from Vector Laboratories (Burlingame, CA, USA). A dual luminescence enzyme kit for luminescent reporter assay was purchased from Promega (Madison, Wis., USA). Super 19X TOPFlash (STF) and 8XFOPFlash (SFF) vectors and Renilla luciferase vectors, Wnt3A-secreted L 292 cells used for luminescent reporter analysis were obtained from Amiee Kohn's laboratory.

연구방법Research method

1. 세포배양1. Cell culture

갑상선암 세포주들은 각 세포에 적합한 배지를 이용하여 5% 이산화탄소, 37℃ 조건의 항온배양기에서 배양되었다. 대표적으로 이용된 BCPAP와 TPC-1 세포주는 10% 우태아 혈청과 항생제를 첨가한RPMI-1640 배지에서 배양되었고, Cal62 세포주는 20% 우태아 혈청과 항생제를 첨가한 DMEM 배지에서 배양되었다. 배양액은 2-3일마다 교환하였으며 세포가 80% 정도 차면 계대배양을 하였다.
Thyroid cancer cell lines were cultured in 5% CO2 and 37 ° C incubator using media suitable for each cell. BPCAP and TPC-1 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and antibiotics. Cal62 cells were cultured in DMEM supplemented with 20% fetal bovine serum and antibiotics. The culture medium was changed every 2-3 days and subculture was performed when the cells reached about 80%.

2. 실시간 중합 효소 연쇄 반응 2. Real-time polymerase chain reaction

세포에서 RNA 분리는 RNeasy Mini 키트 (Qiagen) 를 이용하였는데, 세포의 펠렛에 600 의 RLT 완충용액과 beta-mercaptoethanol 혼합용액을 넣고 섞어준 후 70% 에탄올 동량을 넣고 10000 rpm에서 1분간 원심분리한다. 하등액은 버리고 RWI 용액 700 , RPE 500 , RPE 500 용액을 차례로 넣고 원심분리 하면서 하등액을 버리고 말린다. RNase free 증류수 30 를 넣고 5분간 기다린 후 다시 13000 rpm에서 2분간 원심분리하고 얻어진 하등액으로 RNA 정량하였다. For RNA isolation from the cells, RNeasy Mini Kit (Qiagen) was used. To the cell pellet, 600 RLT buffer solution and beta-mercaptoethanol solution were added and mixed. 70% ethanol was added and centrifuged at 10000 rpm for 1 minute. Discard the lower solution, add RWI solution 700, RPE 500, and RPE 500 solution in this order, centrifuge, discard the lower solution, and dry. RNase free distilled water 30 was added and the mixture was centrifuged at 13000 rpm for 5 minutes. RNA was quantified by the obtained solution.

조직에서 RNA 분리는 TRIzol RNA 분리시약을 이용하였고 조직에 1 mL의 trizol 용액을 투여한 후 갈고, 클로로포름 200 을 넣고 3분간 섞은 후 4℃, 12000 rpm에서 15분간 원심분리 하였다. 상층액만 모아 이소프로필 알콜 500 을 추가하여 섞어주고 이 후 10분간 상온에서 반응시켰다. 다시 10분간 원심분리 후 상층액을 버리고 75% 에탄올 1 mL을 추가하여 섞어준 후 4℃, 15000 rpm에서 5분간 원심분리하였다. 이 후 상층액을 버리고 말린 후 RNase free 증류수 30 을 넣고 60℃에서 10 여분간 열고정한 후 정량하였다.For RNA isolation, TRIzol RNA isolation reagent was used. 1 mL of trizol solution was added to the tissue, the mixture was shaken, chloroform 200 was added, and the mixture was mixed for 3 minutes and then centrifuged at 4 ° C and 12,000 rpm for 15 minutes. The supernatant was collected, and isopropyl alcohol 500 was added thereto, followed by reaction at room temperature for 10 minutes. After centrifuging again for 10 minutes, the supernatant was discarded and 1 mL of 75% ethanol was added and mixed. The mixture was centrifuged at 15,000 rpm for 5 minutes at 4 ° C. After the supernatant was discarded, dried, and RNase-free distilled water (30) was added.

추출된 RNA 에서 RevertAid 첫가닥 상보적DNA 합성키트 (Fermentas) 를 이용하여 첫 번째 상보DNA를 합성하였다. RNA 1 에 oligo(dT) 18 primer (0.5 / ) 1 와 DEPC 처리수를 추가하여 12 를 만들고 5배 reaction buffer, RioLock RNase 억제제 (20 U/) 1 , 10 mM dNTP 2 , RevertAid 역전사효소 (200 U/) 1 를 추가하여 총 20 를 만든다. 시료를 42℃ 60분, 70℃ 5분간 반응한 후 얼음에 식혔다.The first complementary DNA was synthesized from the extracted RNA using the RevertAid first strand complementary DNA synthesis kit (Fermentas). RNA 1 was added with oligo (dT) 18 primer (0.5 /) 1 and DEPC-treated water to make 12, and 5-fold reaction buffer, 20 L / rat RNase inhibitor, 20 mM dNTP 2, RevertAid reverse transcriptase /) 1 to make a total of 20. The sample was reacted at 42 DEG C for 60 minutes and at 70 DEG C for 5 minutes and then cooled on ice.

이 후 QuantiFastSYBRGreen PCR 키트를 이용하였고 합성된 상보 DNA 1 에 2배 QuantiFast SYBR Green PCR 혼합액 12.5 와 10 pM의 전방 및 후방 프라이머 각각1 , RNAse free 증류수를 혼합하여 25 의 시료를 만들었다. 사용된 프라이머 염기 서열과 생성물 크기는 다음과 같다. Robo1, 전방 프라이머 5’-ccatcagtccactgccactc-3’, 후방 프라이머 5’-catgtggccagtctcctctg-3’, 92 bp; Robo2, 전방 프라이머 5’-agtgtcaccttgtcctggca-3’, 후방 프라이머 5’-gccagctgttgctcactgat-3’, 101 bp; Robo3, 전방 프라이머 5’-ctttggattgggcatgtgag-3’, 후방 프라이머 5’-actgagggagccagatgctt-3’, 100 bp; Robo4, 전방 프라이머 5’-gtttcggtctagagcccagg-3’, 후방 프라이머 5’-aggaagatctcatcccgtgg-3’, 86 bp ; SLIT1 , 전방 프라이머 5’-gtgtgggcaactacacctgc-3’, 후방 프라이머 5’-atgggttcagatccggagag-3’, 97 bp ; SLIT2, 전방 프라이머 5’-gcttgccaaacaaccaagaa-3’, 후방 프라이머 5’-acaaaggaggagccgtcagt-3’, 136 bp; SLIT3, 전방 프라이머 5’- gcagaagcccatagcaacaa-3’, 후방 프라이머 5’-ttccatttgttcgaccctca-3’, 126 bp. 18S rRNA가 housekeeping gene으로 이용되었고 이의 전방 프라이머는 5’-aaacggctaccacatccaag-3’, 후방 프라이머는5’-cctccaatggatcctcgtta-3’ 였다. PCR에는 7500 Fast Real-time PCR system (Applied biosystems, Foster city, CA, USA) 이 사용되었고 95℃에서 10초간 분리, 60℃에서 30초간 합성되는 과정을 40주기 반복하여 증폭하였다. 이 후 증폭 곡선을 그려 정량하였다.
After that, QuantiFastSYBRGreen PCR kit was used. Twenty-five specimens were prepared by mixing 2x QuantiFast SYBR Green PCR mixture 12.5, 10 pM each of forward and backward primer 1, and RNAse free distilled water to synthesized complementary DNA 1. The primer sequences and product sizes used were as follows. Robo1, forward primer 5'-ccatcagtccactgccactc-3 ', back primer 5'-catgtggccagtctcctctg-3', 92 bp; Robo2, forward primer 5'-agtgtcaccttgtcctggca-3 ', back primer 5'-gccagctgttgctcactgat-3', 101 bp; Robo3, forward primer 5'-ctttggattgggcatgtgag-3 ', rear primer 5'-actgagggagccagatgctt-3', 100 bp; Robo4, forward primer 5'-gtttcggtctagagcccagg-3 ', back primer 5'-aggaagatctcatcccgtgg-3', 86 bp; SLIT1, forward primer 5'-gtgtgggcaactacacctgc-3 ', back primer 5'-atgggttcagatccggagag-3', 97 bp; SLIT2, forward primer 5'-gcttgccaaacaaccaagaa-3 ', back primer 5'-acaaaggaggagccgtcagt-3', 136 bp; SLIT3, forward primer 5'-gcagaagcccatagcaacaa-3 ', back primer 5'-ttccatttgttcgaccctca-3', 126 bp. 18S rRNA was used as a housekeeping gene and its forward primer was 5'-aaacggctaccacatccaag-3 'and its rear primer was 5'-cctccaatggatcctcgtta-3'. PCR was performed using a 7500 Fast Real-time PCR system (Applied Biosystems, Foster City, Calif., USA), which was amplified by repeating 40 cycles of 95 ° C for 10 seconds and 60 ° C for 30 seconds. After that, an amplification curve was drawn and quantified.

3. 갑상선 조직 코호트 구성 및 조직미세배열 슬라이드제작3. Thyroid tissue cohort and tissue microarray slides

본 연구는 본원의 임상연구심의위원회의 허가를 얻어 2003년부터 2012년까지 본원에서 갑상선 유두암 또는 양성 갑상선 종양으로 갑상선 절제술을 시행 받고 수술 후 보관 중인 파리핀 블록 조직이 있는 239명의 환자들로 갑상선암 조직 코호트를 구성하였고 이들의 임상병리학적 소견에 대한 자료를 수집하였다. 이 중 양성 갑상선 결절인 결절성 과증식증과 여포성 선종 조직을 보이는 경우는 각각 36명, 41명이었고 갑상선 유두암 환자가 160명 이었는데 이들 중 대응되는 정상 갑상선 조직이 있는 환자는 154명이었고 림프절 전이를 동반한 경우는 68명 이었다. 전체 393개의 조직에 대해 조직미세배열 슬라이드를 제작하였다. The purpose of this study was to evaluate the clinical characteristics of thyroid carcinoma tissues in patients with thyroid papillary carcinoma or benign thyroid carcinoma. The cohort was constructed and the data of clinicopathological findings were collected. Of these, 36 had nodular hyperplasia and follicular adenoma, 41 had papillary thyroid nodules, and 160 had papillary thyroid cancer. Among them, 154 patients had normal thyroid tissue, and lymph node metastasis One case was 68 patients. Tissue microarray slides were prepared for a total of 393 tissues.

모든 조직은 Hematoxylin-Eosin 염색을 시행하였고 숙련된 병리 의사에 의해 다시 한 번 검토 후 적절한 부분을 선택하여 조직미세배열 슬라이드를 제작하였다. 슬라이드 제작에는 tissue-arraying instrument (Beecher Instruments, Sliver Spring, USA)가 사용되었으며 공여 블록에서 지름 2 mm의 조직 실린더를 이용하여 지정한 부분을 펀치하여 수여 블록으로 옮겼고 정상 조직과 양성 갑상선 종양 조직에서는 한 샘플당 1개, 갑상선 유두암 조직에서는 한 샘플당 2개의 코어를 얻었다.
Hematoxylin-Eosin staining was performed on all tissues, and a tissue microarray slide was prepared by a skilled pathologist after reviewing the appropriate sections. A tissue-arraying instrument (Beecher Instruments, Sliver Spring, USA) was used for the preparation of the slides. The specimens were transferred from the donor block to a given block using a 2 mm diameter tissue cylinder. In normal tissue and benign thyroid tumors, one sample One per sample, and two cores per sample in papillary thyroid tissue.

4. 갑상선 조직의 면역화학염색4. Immunochemical staining of thyroid tissue

조직미세배열 슬라이드를 이용하여 SLIT2 (Santa Cruz Biotechnology, Santa Cruz, USA)와 Robo1(Abcam, Cambridge, UK) 에 대한 면역화학염색을 시행하였고 Bench markautomatic immunostaining device (Roche Tissue Diagnostics, Tucson, USA)와 UltraviewTM Universal DAB Detection 키트(Ventana Medical Systems, Tucson, USA)가 이용되었다. 면역화학염색 결과는 반정량적으로 양성 또는 음성으로 판단되었는데, 숙련된 병리과 의사가 염색 강도를 0-3점으로 분류하고 2점 이상인 경우를 양성으로 하였다. 대표적인 예시를 도 2 및 도 3에 정리하였다.
Immunohistochemical staining was performed with SLIT2 (Santa Cruz Biotechnology, Santa Cruz, USA) and Robo1 (Abcam, Cambridge, UK) using tissue microarray slides. Universal DAB Detection kit (Ventana Medical Systems, Tucson, USA) was used. Immunohistochemical stainings were judged to be positive or negative semi-quantitatively. A skilled pathologist graded the staining intensity as 0-3 and positive as 2 or more. Representative examples are summarized in FIG. 2 and FIG.

5. 갑상선 조직 BRAF V600E 돌연변이 분석5. Thyroid tissue BRAF V600E mutation analysis

SLIT-Robo 경로의 발현과 BRAF 돌연변이의 연관성을 확인하기 위해 조직미세배열 슬라이드를 제작한 갑상선 조직 코호트에서 DNA를 추출하여 BRAF V600E변이 유무를 확인하였다. 파라핀블록은 미세절단하였고, 미세절단된 조직에서 QIAampDNA FFPE 조직키트를 이용하여 유전체 DNA를 추출하였다. V600E 변이가 위치한 BRAF 유전자의 exon 15번을 증폭하였고 전방 프라이머는 5’-tgcttgctctgataggaaaa-3’, 후방 프라이머는 5’-ctgatgggacccactccat-3’을 이용하였으며 Takara PCR thermal cycler (Applied biosystems) 에서 94℃ 에서 30초, 58℃에서 30초, 70℃에서 30초의 과정을 36회 반복하였다. 증폭된 DNA는 1% 한천 겔에서 전기 영동하였으며 Gel-doc 시스템에서 사진을 찍어 확인하였다.
To confirm the association between the expression of the SLIT-Robo pathway and the BRAF mutation, DNA was extracted from the thyroid tissue cohort in which the tissue microarray slides were prepared and the presence of BRAF V600E mutation was confirmed. The paraffin block was microfuncated and genomic DNA was extracted from the microtitered tissue using a QIAamp DNA FFPE tissue kit. The exon 15 of the BRAF gene in which the V600E mutation was located was amplified using 5'-tgcttgctctgataggaaaa-3 'as the forward primer and 5'-ctgatgggacccactccat-3' as the back primer. The PCR reaction was carried out using a Takara PCR thermal cycler (Applied biosystems) Sec, 58 [deg.] C for 30 sec, and 70 [deg.] C for 30 sec were repeated 36 times. The amplified DNA was electrophoresed on 1% agar gel and photographed on a Gel-doc system.

6. 세포 생존 및 이동능 분석6. Cell survival and migration ability analysis

세포 생존은 직접 세포 수를 측정하는 방법과 Bromodeoxyuridine(BrdU)을 이용한 발색분석법을 이용하여 확인하였다. 세포 수는 24 웰 플레이트에 각각 1.5X10³ 세포를 분주하고 24시간 후 다양한 농도의 SLIT을 투여하고 72시간 후 측정하였는데, 10 의 시료와 동량의 트립판 블루 용액을 혼합한 후 EVETM automated cell counter (NanoEnTek, Korea)를 이용하였다. BrdU 발색분석은 96웰 플레이트에 3000개의 세포를 100 로 분주한 후 24시간 후 다양한 농도의 SLIT을 투여하고 48시간 후 진행하였고, 10 의 BrdU labeling 용액을 각 웰 플레이트에 분주하고 2시간 반응시킨 후 배양액을 제거하고 200 의 fixDent 용액을 넣고 30분간 상온에서 반응시킨 후 이를 제거하고 100 의 anti-BrdU POD 용액을 넣고 90분간 반응, 세척 후 기질 용액을 넣고 370 nm 파장에서 흡광도를 측정하였다.Cell viability was confirmed by direct cell count and colorimetric assay using Bromodeoxyuridine (BrdU). Cells were plated in 24-well plates at 1.5 × 10 ³ cells, and after 24 hours, various concentrations of SLIT were administered. After 72 hours, 10 samples were mixed with the same amount of tryptamine blue solution, followed by EVETM automated cell counter NanoEnTek, Korea) was used. BrdU color analysis was performed by dividing 3000 cells into 100 wells of a 96-well plate. After 24 hours, various concentrations of SLIT were administered. Forty-eight hours later, 10 BrdU labeling solutions were dispensed into each well plate and reacted for 2 hours After incubation for 30 min at room temperature, 100 of anti-BrdU POD solution was added. After incubation for 90 min, substrate solution was added and absorbance was measured at 370 nm wavelength.

세포 이동능은 상처치유분석 및 transwell chamber 침습검사를 통해 측정하였는데, 상처치유분석은 세포를 6 웰 플레이트에 8 X 10⁵ 개 씩 분주하고 24시간 후 상처를 만들고 3 nM의 Slit2 와 Slit3 을 처리한 후 24시간 동안의 상처거리 변화를 관찰하였다. Transwell chamber 침습검사는 CytoSelect TM 24-well cell invasion assay 키트를 이용하여 내부 공간에는 세포 5 X 10⁵ 개와 우태아혈청 무첨가 배지 및 Slit2 3 nM 및 Slit3 3 nM를 넣었고, 외부 공간에는 우태아혈청 첨가 배지를 넣고 매뉴얼에 따라 실험을 진행하였다. 결과는 2 곳의 고배율(X 1000) 시야 에서 염색된 세포 수를 측정하여 평균값을 구하고 추출 용액으로 염색약을 녹여낸 후 560 nm 파장에서 흡광도를 측정한 것을 나타내었다.
Cell migration was measured by wound healing analysis and transwell chamber invasion assay. The wound healing analysis was performed by dividing the cells into 6 well plates at 8 × 10 ⁵ , and after 24 hours, the wound was prepared and treated with 3 nM of Slit2 and Slit3 The wound changes were observed for 24 hours. The transwell chamber invasion test was performed using a CytoSelect ™ 24-well cell invasion assay kit, in which 5 × 10 ⁵ cells, 5% fetal bovine serum-free medium, 3 nM Slit2 and 3 nM Slit3 were added. And the experiment was conducted according to the manual. The results were obtained by measuring the number of cells stained in two high power fields (X 1000), measuring the absorbance at 560 nm after dissolving the dye in the extract solution.

7. 웨스턴 블롯 분석7. Western blot analysis

단백 추출을 위해 샘플을 만들 세포를 차게 유지된 PBS 용액으로 2회 세척하여 반응을 종료시켰다. 이 후 단백질 분해 효소 억제제를 첨가한 세포용해완충액을 이용하여 세포를 모아 분해한 후 4℃, 13,000 rpm에서 30분 동안 원심분리하였고, 단백질을 포함하는 상층액을 분리하여 단백질 정량을 하였다. 30 ug의 단백질을 10% NuPAGE 겔에서 전기영동하여 전개시킨 후 니트로셀룰로스막으로 이동시켰다. 막에 해당하는 일차항체 (1:1000 희석)와 18시간 동안 냉장 반응 시킨 후, HRP가 연결되어 있는 이차항체 (1:5000 희석)로 2시간 동안 반응 시켰으며 이 후HRP 화학발광면역기질시약을 이용하여 필름 감광 하였다. 단백 발현량은 각각의 샘플에서 GAPDH 발현량으로 비교하였다.
The cells to be sampled for protein extraction were washed twice with cold PBS solution to terminate the reaction. Then, the cells were collected using a cell lysis buffer containing protease inhibitor, and then the cells were centrifuged at 13,000 rpm for 30 minutes at 4 ° C. Protein-containing supernatants were separated and quantified. 30 ug of protein was developed by electrophoresis in 10% NuPAGE gel and transferred to nitrocellulose membrane. (1: 1000 dilution) for 18 hours and then reacted with secondary antibody (1: 5000 dilution) with HRP for 2 hours. After that, HRP chemiluminescent immune substrate reagent The film was photosensitive. Protein expression levels were compared by GAPDH expression in each sample.

8. 발광 리포터 분석8. Analysis of luminescent reporter

24웰 플레이트에 1 X 10⁵ 세포를 분주하고 70% 정도 차면 DNA 형질주입을 진행하였다. 먼저 각 우물 당 50 의 DNA-Lipofectamine 혼합액 (500 ng의 STF 또는 SFF 및 50 ng renilla 벡터, lipofectaminTM 2000 1.5 ) 을 넣고 4시간 동안 반응시켰다. 이 후 각 우물의 조건에 따라 Wnt3A 조건배지 또는 일반 배지를 투여하였고, SLIT 투여가 필요한 경우 SLIT2 5 nM을 투여하였다. Wnt3A 조건 배지는 L292 세포를 안정화되게 키운 후 세포가 100 mm 배양 접시에서 90% 정도 찼을 때2% 우태아혈청을 포함한 DMEM 배지를 넣어 2-3일 째에 배지를 모아 0.45 로 필터하여 사용하였고, 세포에는 20% 농도로 투여하였다. 24시간 후 발광효소 활성도를 확인하였고, 먼저 100 발광효소분석 시약과 20 세포 lysate를 반응시킨 후 firefly 발광 활성을 확인, Stop & Go 용액 100 를 투여한 후 renilla 발광효소 발광활성을 확인하였으며, 결과는 firefly 발광 활성도를 renilla 발광 활성도로 나눈 값으로 나타내었다. 1 × 10 ⁵ cells were dispensed into a 24-well plate and the DNA transfection was carried out when the cell concentration reached about 70%. First, 50 DNA-Lipofectamine mixtures (500 ng STF or SFF and 50 ng renilla vector, lipofectamin 占 2000 1.5) were added per well and reacted for 4 hours. After this, Wnt3A condition medium or normal medium was administered according to the condition of each well, and SLIT2 5 nM was administered when SLIT administration was necessary. Wnt3A conditioned medium was prepared by stabilizing L292 cells. When the cells were 90% full in a 100 mm culture dish, DMEM medium containing 2% fetal bovine serum was added. After 2-3 days, the medium was collected and filtered to 0.45, The cells were administered at a concentration of 20%. After 24 hours, the activity of the luminescent enzyme was confirmed. First, 100 luminescent enzyme assay reagent and 20 cell lysate were reacted, and firefly luminescence activity was confirmed. After the Stop & Go solution 100 was administered, The firefly luminescence activity was expressed as the value divided by the renilla luminescence activity.

이 실험은 Wnt/beta-catenin 경로의 활성도를 알아보기 위한 방법으로 Wnt가 세포 표면 복합 수용체와 결합하면 이것은 beta-catenin의 인산화를 억제, beta-catenin의 핵 내로의 이동을 촉진하고 이는 lymphoid enhancer factor/T-cell factor (LEF/TCF) 전사 인자와 결합하여 유전자 발현을 조절하는 것을 이용하였다. STF는 정상적인 LEF/TCF 전사인자와 firefly 발광효소가 결합된 벡터이고, SFF는 변형된 LEF/TCF 전사인자가 결합된 것으로 음성 대조군으로 이용된다.²⁹⁾
When Wnt binds to the cell surface complex, it inhibits the phosphorylation of beta-catenin and promotes the translocation of beta-catenin into the nucleus, which results in the activation of the lymphoid enhancer factor / T-cell factor (LEF / TCF) transcription factor to regulate gene expression. STF is a vector with normal LEF / TCF transcription factor and firefly luciferase, and SFF is a negative control with modified LEF / TCF transcription factor. ²⁹⁾

결과result

1. 갑상선암 세포주와 갑상선암 조직의 SLIT-Robo mRNA 발현1. SLIT-Robo mRNA expression of thyroid cancer cell line and thyroid cancer tissue

다양한 갑상선암 세포주에서 실시간 중합효소 연쇄반응을 통해 SLIT과 Robo 아형의 mRNA 발현량을 확인하였다. 그림에서 보는 것과 같이 갑상선암 모든 세포주에서 모든 SLIT과 Robo 아형의 mRNA 발현이 확인되었는데, 특히 Robo1, Robo4 및 Slit2, Slit3의 발현이 많았다(도 4a 및 도 4b). 신선 동결 갑상선암 조직과 그에 대응되는 정상 갑상선 조직에서 12 개에서 Slit과 Robo의 발현을 확인하였을 때도 정상 조직에 비해 갑상선암 조직에서 SLIT과 Robo 모든 아형의 발현이 증가하였다 (도 5).
The mRNA expression levels of SLIT and Robo subtypes were confirmed by real - time PCR in various thyroid cancer cell lines. As shown in the figure, mRNA expression of all SLIT and Robo subtypes was confirmed in all cell lines of the thyroid cancer, especially Robo1, Robo4, Slit2 and Slit3 (FIGS. 4A and 4B). When SLIT and Robo were expressed in 12 frozen thyroid carcinoma tissues and corresponding thyroid tissues, the expression of SLIT and Robo all subtypes was increased in thyroid cancer tissues compared with normal tissues (Fig. 5).

2. 갑상선 조직의 Slit-Robo 발현2. Slit-Robo expression in thyroid tissue

갑상선암 조직 코호트를 대상으로 제작한 조직 미세배열 슬라이드를 이용하여 Robo1 및 SLIT2의 면역화학염색을 시행하였다. Robo1의 경우 정상 조직, 양성 갑상선 종양에서는 각각 29%, 21% 의 양성률을 보였으나 갑상선 유두암 조직에서는 81%에서 양성을 보여 정상 및 양성 조직 보다 갑상선 유두암 조직에서 발현이 유의하게 증가하였다. Slit2 또한 유사한 결과를 보였는데, 양성 갑상선 종양에서 Slit2의 양성률은 65%, 갑상선 유두암 조직의 양성률은 85%였고, 정상 조직에서는 2.6%로 정상 또는 양성 갑상선 조직에 비해 갑상선 유두암 조직의 Slit2 발현이 유의하게 증가하는 것을 확인하였다. (도 6 및 도 7).
Immunohistochemical staining of Robo1 and SLIT2 was performed using tissue microarray slides prepared for thyroid carcinoma tissue cohort. In the case of Robo1, 29% and 21% of normal and positive thyroid tumors were positive, respectively, but in 81% of cases of papillary thyroid carcinoma tissues, expression was significantly increased in papillary thyroid tissue tissues than normal and positive tissues. Slit2 also showed similar results. The positive rate of Slit2 was 65% in the positive thyroid tumor, 85% in the pituitary tumor tissue, and 2.6% in the normal tissue, showing the Slit2 expression of the papillary thyroid tissue compared with the normal or positive thyroid tissue . (Figs. 6 and 7).

3. 면역화학염색에 따른 Slit-Robo 발현 정도와 환자 임상양상 비교 분석3. Comparison of Slit-Robo expression level and clinical outcome according to immunochemical staining

조직 미세배열 슬라이드 면역화학염색 결과에 따른 Slit-Robo 발현 정도와 환자의 임상 양상을 비교 분석하였다. Slit2와 Robo1의 발현 저하는 림프절 전이의 증가와 통계적으로 유의한 관련을 보였는데 림프절 전이가 없는 갑상선 유두암의 Robo1 양성률은 87%, Slit2 양성률을 95% 이나 림프절 전이가 동반된 경우 각각 72%, 72% 로 낮았다. 특히 Slit2 발현 감소는 림프절 전이뿐 만 아니라 종양의 림프혈관계 침범, 원격전이 및 재발 위험 증가와도 통계적으로 유의한 연관을 보였다. 그러나 이러한 연관성은 BRAF 변이 유무와는 무관하였다. (도 8, 표 1, 표 2, 그림 도 9)Slit-Robo expression and clinical characteristics of patients were compared according to the result of immunohistochemical staining. There was a statistically significant correlation between the decreased expression of Slit2 and Robo1 in the lymph node metastasis. Robo1 and Slit2 positivity was 87% and 72% in 72 and 72% of cases, respectively. %. In particular, the decreased expression of Slit2 was associated with not only lymph node metastasis, but also increased risk of lymphoma involvement, distant metastasis and recurrence. However, this association was not related to the presence or absence of BRAF mutation. (Fig. 8, Table 1, Table 2, Fig. 9)

Robo1 발현에 따른 임상양상 비교Comparison of clinical features according to Robo1 expression -- 전체(total)Total (total) Robo1Robo1 p-value p-value 음성voice 양성positivity numbernumber 160160 3131 129129 -- 나이age 47.4 ± 13.647.4 ± 13.6 46.0 ± 13.046.0 + - 13.0 47.7 ± 13.847.7 ± 13.8 0.510.51 성 (male)Sex (male) 30 (18.8%)30 (18.8%) 4 (12.9%)4 (12.9%) 26 (20.2%)26 (20.2%) 0.350.35 종양크기 (cm)Tumor size (cm) 2.6 ± 1.32.6 ± 1.3 2.9 ± 1.42.9 ± 1.4 2.5 ± 1.32.5 ± 1.3 0.080.08 갑상선외 침윤External thyroid infiltration 119 (74.4%)119 (74.4%) 20 (64.5%)20 (64.5%) 99 (76.7%)99 (76.7%) 0.160.16 림프혈관 침윤Lymphovascular infiltration 10 (6.3%)10 (6.3%) 4 (12.9%)4 (12.9%) 6 (4.7%)6 (4.7%) 0.090.09 자궁경부 림프절 전이Cervical lymph node metastasis 68 (42.5%)68 (42.5%) 19 (61.3%)19 (61.3%) 49 (38.0%)49 (38.0%) 0.0250.025 원발성 전이Primary metastasis 17 (10.6%)17 (10.6%) 5 (16.1%)5 (16.1%) 13 (10.1%)13 (10.1%) 0.340.34 재발율Recurrence rate 36/155 (23.2%)36/155 (23.2%) 5 (16.7%)5 (16.7%) 31 (24.8%)31 (24.8%) 0.470.47 BRAFBRAF ^V600EV600E 113/158113/158 21 (70.0%)21 (70.0%) 92 (71.9%)92 (71.9%) 0.840.84

Slit2 발현에 따른 임상양상 비교Comparison of clinical features according to Slit2 expression -- 전체(total)Total (total) SLIT2SLIT2 p-value p-value 음성voice 양성positivity numbernumber 160160 2424 136 136 -- 나이age 47.4 ± 13.647.4 ± 13.6 44.7 ± 13.144.7 ± 13.1 47.9 ± 13.747.9 ± 13.7 0.290.29 성 (male)Sex (male) 30 (18.8%)30 (18.8%) 7 (29.2%)7 (29.2%) 23 (16.9%)23 (16.9%) 0.160.16 종양크기 (cm)Tumor size (cm) 2.6 ± 1.32.6 ± 1.3 2.9 ± 1.52.9 ± 1.5 2.5 ± 1.3
2.5 ± 1.3
0.20 0.20 갑상선외 침윤External thyroid infiltration 119 (74.4%)119 (74.4%) 18 (75.0%)18 (75.0%) 101 (74.3%)101 (74.3%) 0.940.94 림프혈관 침윤Lymphovascular infiltration 10 (6.3%)10 (6.3%) 4 (16.7%)4 (16.7%) 6 (4.4%)
6 (4.4%)
0.0220.022 자궁경부 림프절 전이Cervical lymph node metastasis 68 (42.5%)68 (42.5%) 19 (79.2%)19 (79.2%) 49 (36.0%)49 (36.0%) < 0.001<0.001 원발성 전이Primary metastasis 17 (10.6%)17 (10.6%) 10 (41.7%)10 (41.7%) 8 (5.9%)8 (5.9%) < 0.001<0.001 재발율Recurrence rate 36/155 (23.2%)36/155 (23.2%) 14 (63.6%)14 (63.6%) 22 (16.5%)22 (16.5%) < 0.001<0.001 BRAFBRAF ^V600EV600E 113/158113/158 14 (58.3%)14 (58.3%) 99 (73.9%)99 (73.9%) 0.12 0.12

또한, 대상 환자들을 Slit2와 Robo1의 발현이 모두 양성인 군과 그렇지 않은 군으로 나누어 추가 분석을 시행하였을 때, Slit2와 Robo1이 모두 양성인 경우 종양의 크기가 더 작고 림프절 전이 및 원격전이, 재발이 통계적으로 유의하게 적게 발생함을 확인할 수 있었다 (표 3)In addition, when Slit2 and Robo1 were both positive, both Slit2 and Robo1 were positive. When both Slit2 and Robo1 were positive, tumor size was smaller and lymph node metastasis, distant metastasis and relapse were statistically significant (Table 3)

Robo1 및 Slit2 발현에 따른 임상양상 비교Comparison of clinical features according to Robo1 and Slit2 expression -- 전체(total)Total (total) Robo1 & SLIT2Robo1 & SLIT2 p-value p-value 기타(others)Others 모두 양성All positive numbernumber 160160 5454 106106 -- 나이age 47.4 ± 13.647.4 ± 13.6 45.7 ± 13.645.7 ± 13.6 48.3 ± 13.648.3 ± 13.6 0.260.26 성 (male)Sex (male) 30 (18.8%)30 (18.8%) 10 (18.5%)10 (18.5%) 20 (18.9%)
20 (18.9%)
0.960.96 종양크기 (cm)Tumor size (cm) 2.6 ± 1.32.6 ± 1.3 3.0 ± 1.73.0 ± 1.7 2.4 ± 1.02.4 ± 1.0 0.0060.006 갑상선외 침윤External thyroid infiltration 119 (74.4%)119 (74.4%) 35 (64.8%)35 (64.8%) 84 (79.2%)84 (79.2%) 0.060.06 림프혈관 침윤Lymphovascular infiltration 10 (6.3%)10 (6.3%) 6 (11.1%)6 (11.1%) 4 (3.8%)4 (3.8%) 0.090.09 자궁경부 림프절 전이Cervical lymph node metastasis 68 (42.5%)68 (42.5%) 35 (64.8%)35 (64.8%) 33 (31.1%)33 (31.1%) < 0.001<0.001 원발성 전이Primary metastasis 17 (10.6%)17 (10.6%) 12 (22.2%)12 (22.2%) 6 (5.7%)6 (5.7%) 0.0030.003 재발율Recurrence rate 36/155 (23.2%)36/155 (23.2%) 17 (33.3%)17 (33.3%) 19 (18.3%)19 (18.3%) 0.0440.044 BRAFBRAF ^V600EV600E 113/158113/158 34 (64.2%)34 (64.2%) 79 (75.2%)79 (75.2%) 0.1910.191

4. Slit 투여에 따른 갑상선암 세포주 증식 변화4. Proliferation of thyroid cancer cell line by administration of Slit

이러한 조직 면역염색결과를 바탕으로 본 연구자들은 갑상선암 세포 모델에서 Slit 투여에 따른 갑상선암 세포주의 변화를 관찰하기로 하였다. 먼저 Slit 투여에 따른 갑상선암 세포주의 증식 변화를 확인하기 위해 무작위로 3가지 갑상선암 세포주 (BCPAP, TPC-1, Cal62)를 선택하였고 0.1 에서 10 nM 사이의 다양한 농도의 Slit2 와 Slit3를 투여하고 72시간 후 세포 성장 변화를 확인하였다. 세포 성장 변화는 세포 수 측정 및 BrdU 측정의 두 가지 방법으로 확인하였으며, 두 방법은 유사한 경향을 보였는데, Slit2 와 Slit3 모두 비교적 농도 의존적으로 세포의 성장을 억제하였다. 그 효과는 갑상선 유두암 유래 세포주인 BCPAP 및 TPC-1에서 좀 더 뚜렷하였는데, 10 nM 농도의 Slit2 투여 시 BCPAP는 최대 43%, TPC-1 는 33% 의 성장 억제 효과가 관찰되었다 (도 10 및 도 11).
Based on these immunohistochemical stainings, we decided to observe the changes of thyroid cancer cell line according to Slit administration in the thyroid cancer cell model. Three thyroid carcinoma cell lines (BCPAP, TPC-1, Cal62) were selected randomly, and Slit2 and Slit3 at various concentrations ranging from 0.1 to 10 nM were administered to examine the proliferation of thyroid cancer cell line according to Slit administration. Cell growth changes were confirmed. Cell growth was determined by two methods: cell counting and BrdU assay. Both methods showed a similar tendency. Both Slit2 and Slit3 inhibited cell growth in a relatively dose - dependent manner. The effect was more pronounced in the papillary thyroid cancer cell lines BCPAP and TPC-1. In Slit2 administration of 10 nM, BCPAP up to 43% and TPC-1 33% growth inhibitory effect were observed (FIGS. 10 and 11).

5. Slit 투여에 따른 갑상선암 세포주 이동능 변화5. Changes in thyroid carcinoma cell migration by Slit administration

BCPAP 갑상선암 세포주를 선택하여 상처치유분석을 통해 Slit 투여에 따른 갑상선암 세포주 이동능 변화를 확인하였다. 3 nM 농도의 Slit2 와 Slit3를 투여하고 24시간 동안 관찰하였을 때 Slit2 및 Slit3 모두 상처치유를 뚜렷하게 억제함을 확인할 수 있었다 (도 12). 또한 Transwell chamber 침습분석에서도 Slit2 와 Slit3 3 nM 투여에 의한 세포 이동능 억제 효과를 관찰할 수 있었다 (도 13).
BCPAP thyroid cancer cell line was selected and the change of thyroid carcinoma cell migration ability by Slit administration was confirmed by wound healing analysis. When Slit2 and Slit3 were administered at a concentration of 3 nM and observed for 24 hours, both Slit2 and Slit3 significantly inhibited wound healing (Fig. 12). In addition, the inhibition of cell migration by Slit2 and 3 nM of Slit3 was also observed in the Transwell chamber invasion assay (Fig. 13).

6. Slit 투여가 세포 성장과 이동에 관련된 신호 전달체계에 미치는 영향6. Effects of Slit administration on signaling pathways involved in cell growth and migration

Slit 투여로 성장 억제 및 뚜렷한 세포 이동 감소를 보인 BCPAP 세포주에서 웨스턴 블롯 분석을 통해 세포 내 신호 전달 체계의 변화를 확인하였다. 기존 연구에서 알려진 바와 같이 갑상선암 세포주에서도 Slit 투여는 활성화 beta-catenin의 발현을 억제하고 E-cadherin 활성을 유도하였다 (도 14).
Western blot analysis of the BCPAP cell line showing growth inhibition and marked cell migration reduction by Slit administration confirmed changes in the intracellular signaling pathway. As previously reported, Slit administration also inhibited the expression of activated beta-catenin and induced E-cadherin activity in thyroid cancer cell lines (Fig. 14).

7. Slit 투여에 따른 wnt3A 유도 beta-catenin 경로 활성 변화7. Effect of Slit on wnt3A induced beta-catenin pathway activity

Slit의 성장억제 및 세포이동능 억제 효과가 beta-catenin의 활성도 저하에 따른 것임을 한 번 더 확인하기 위해 BCPAP 세포주를 이용하여 발광효소 리포터 분석을 시행하였다. STF 가 형질전환된 세포에서는 wnt3A 조건배지를 투여하였을 때 발광효소 활성비가 280 배 가량 증가하였으나 SFF 형질전환된 세포에서는 그러한 증가가 보이지 않았다. Slit2의 투여는 wnt3A 투여로 인해 활성화된 wnt/beta-catenin 경로를 의미 있게 억제하였으며 STF가 형질전환된 세포에서 wnt3A 조건배지와 Slit2를 같이 투여하였을 때 발광효소 활성비는 평균 67배 였다 (도 15).
In order to further confirm that the inhibition of slit growth and the inhibition of cell migration were due to the decreased activity of beta-catenin, luminescent enzyme reporter analysis was performed using BCPAP cell line. In the cells transformed with STF, the luminescence enzyme activity ratio was increased about 280 times by the wnt3A conditioned medium, but such increase was not observed in SFF transformed cells. The administration of Slit2 significantly inhibited the wnt / beta-catenin pathway activated by wnt3A administration, and when the wnt3A conditioned medium and Slit2 were co-administered with STF-transformed cells, the lysozyme activity ratio averaged 67 times (Figure 15 ).

고찰Review

본 연구에서 Slit-Robo 경로의 발현 감소는 림프절 전이, 원격전이, 재발 등 갑상선암의 나쁜 예후와 연관이 있었고, 세포 실험에서 갑상선암 세포주에 Slit을 투여하면 세포의 이동능이 현저히 억제되었으며 beta-catenin의 활성이 억제되고 E-cadherin의 발현이 증가되었다. 이에 Slit-Robo 경로의 발현 감소로 세포 이동능이 증가한 것이 전이 등 나쁜 예후 인자들을 나타내는 종양 표현형을 유발한 것으로 생각된다. 또한, Slit2 및 Slit3의 투여는 농도 의존적으로 갑상선암 세포의 증식을 억제하였고, Slit2 및 Robo1의 조직면역화학염색 결과에서 Slit2 및 Robo1이 모두 양성인 경우 그렇지 않은 경우에 비해 종양 크기가 감소하였다. 이를 통해 Slit-Robo 경로의 활성이 갑상선암의 이동능 뿐 만 아니라 성장 또한 억제하였음을 알 수 있었다. 이러한 결과들은 Slit2가 종양 억제 유전자로서의 역할이 있음을 보고한 이전 다른 암종 연구들의 결과와 일치하며, 갑상선암에서도 Slit이 종양 억제 유전자로서 기능이 있음을 처음으로 확인한 것이다. 또한 본 연구 결과는 갑상선암에서 Slit2가 예후를 예측하는 새로운 바이오마커로서의 역할이 있음을 시사하고 Slit2 단백의 투여가 갑상선암의 새로운 표적 치료제로서 작용할 수 있을 가능성도 보여주었다.In the present study, the decrease in expression of Slit-Robo pathway was associated with poor prognosis of thyroid cancer including lymph node metastasis, distant metastasis and recurrence. In the cell study, Slit administration significantly inhibited the migration of the thyroid cancer cell line and the activity of beta-catenin Was suppressed and expression of E-cadherin was increased. Therefore, increased cell migration due to decreased expression of Slit-Robo pathway is thought to induce tumor phenotype, which is associated with poor prognostic factors such as metastasis. In addition, the administration of Slit2 and Slit3 inhibited the proliferation of thyroid cancer cells in a dose - dependent manner. The tumor immunohistochemical staining of Slit2 and Robo1 showed a decrease in tumor size when both Slit2 and Robo1 were positive. This suggests that the activity of the Slit-Robo pathway inhibited not only the migration of thyroid cancer but also the growth. These results are consistent with previous reports of other carcinomas that reported that Slit2 plays a role as a tumor suppressor gene, and it was the first to confirm that Slit functions as a tumor suppressor gene in thyroid cancer. These results suggest that Slit2 plays a role as a new biomarker in predicting the prognosis of thyroid cancer. Slit2 protein may also be a new target treatment for thyroid cancer.

Beta-catenin은 E-cadherin에 의한 세포-세포간의 부착과 EMT를 조절하는 중요한 인자로 알려져 있다. 이전 많은 연구들에서 beta-catenin/E-cadherin 복합체는 Slit과 연관된 세포 이동능 조절에 연관성이 있는 것으로 알려졌고 대표적으로 유방암 세포를 이용한 연구에서도 주변 연조직에서 Slit2의 발현이 증가하면 beta-catenin 경로 억제를 통해 세포의 이동을 억제함을 밝혔다.24) 갑상선암 세포를 이용한 본 연구에서도 웨스턴 블롯 및 발광리포터 분석을 통해 Slit이 beta-catenin 경로 억제를 통해 그 역할을 나타내는 것을 확인하였다. 하지만 Slit-Robo 경로와 beta-catenin 경로가 어떻게 연관이 되는지 그 중간 분자에 대해서는 별로 알려진 바가 없다. 이전 한 연구는 탈유비퀴틴화 효소 USP33이 Robo1의 안정화를 돕고 GSK3b를 활성화하여 beta-catenin을 억제함을 밝힌 바 있으나 이 것만으로 설명은 힘들다.30) 향후 이 중간 경로 분자에 대한 추가 연구가 필요하다. Beta-catenin is known to be an important factor controlling cell-cell adhesion and EMT by E-cadherin. Previous studies have shown that the beta-catenin / E-cadherin complex is involved in the regulation of cell migration associated with Slit. Typically, breast cancer cells have also been shown to inhibit beta-catenin pathway by increasing the expression of Slit2 in the surrounding soft tissue . In the present study using thyroid cancer cells, we confirmed that Slit plays a role in inhibiting beta-catenin pathway through Western blot and luminescent reporter analysis. However, little is known about the intermediate molecule of how the Slit-Robo pathway and the beta-catenin pathway are related. Previous studies have shown that deubiquitin-catalyzing enzyme USP33 helps stabilize Robo1 and activates GSK3b to inhibit beta-catenin, but this is not the only explanation [30]. Further studies on this intermediate pathway molecule are needed .

본 연구에서는 Slit 투여가 갑성선암의 성장도 억제하는 효과를 보였는데, 이전 폐암 세포주 및 췌장암 세포주를 이용한 연구에서는 Slit2 가 종양의 이동능은 억제하지만 성장을 억제하지는 못하였고, 이의 원인은 밝히지 못하였다.^20,25) 종양의 크기과 관련하여서는 Slit2와 Robo4의 상호작용 또한 고려가 필요한데, 이전 연구들에서 Slit2와 Robo4의 결합은GTPase ARF6의 활성을 억제함으로써 혈관 안정성을 증가시키고 VEGF 등은 억제하여 오히려 Slit2-Robo4 경로의 활성은 암 조직의 성장을 촉진시킬 수 있을 것으로 알려졌다²¹ ^,22). 이에 본 연구자들은 추가적으로 조직미세배열 슬라이드를 이용하여Robo4에 대한 면역조직화학검사를 시행하였으나 대부분의 (155/160) 의 갑상선 유두암 조직에서 Robo4가 양성으로 확인되었고 정상 갑상선 조직의 96%에서도 Robo4가 양성으로 나타나 이를 통해 Robo4의 역할을 추론하기는 어려웠다 (Data not shown). 향후 Robo4의 역할에 대해 확인이 필요할 것으로 생각된다. In this study, Slit administration inhibited the growth of giant adenocarcinoma. Slit2 inhibited the migration of tumor but did not inhibit its growth in previous lung and pancreatic cancer cell lines. . ^{In the} previous studies, the combination of Slit2 and Robo4 increased the vascular stability by inhibiting the activity of GTPase ARF6 and inhibited VEGF and the like, resulting in the inhibition of Slit2- The activity of the Robo4 pathway was found to be able to promote the growth of cancer tissues ²¹ ^{, 22)} . We performed immunohistochemistry on Robo4 using tissue microarray slides. However, Robo4 was found to be positive in most (155/160) thyroid papillary tissues and Robo4 was positive in 96% of normal thyroid tissues , Which makes it difficult to deduce the role of Robo4 (Data not shown). The role of Robo4 will be confirmed in the future.

이전 다른 암종 연구 결과에서 정상 조직에 비해 암조직의 Slit-Robo 경로의 활성이 감소하였던 것과 달리¹⁶ ^,20,25) 갑상선암에서는 정상 조직 보다 암 조직에서 Slit-Robo 경로의 발현이 현저하게 증가하였다. 본 연구에서는 정상에 비해 암 조직에서 Slit2의 발현이 증가하는 원인을 밝히지는 못하였으나 Slit 투여가 갑상선암 세포주 모델에서 뚜렷한 종양의 성장과 전이 억제 효과를 보인 것 및 갑상선 조직 코호트에서 Slit-Robo 경로 활성 저하가 나쁜 예후인자와 결부됨을 고려할 때 정상 조직에서 보다 갑상선암에서 Slit-Robo 경로 활성이 증가하는 것이 갑상선암 발생의 원인이 아닌 그 결과일 수 있다. 즉, 다른 알 수 없는 원인 경로의 변이에 의해 갑상선암이 발달하는 것을 Slit-Robo 경로가 억제하기 위해 이차적으로 증가되었다가, 이러한 증가가 억제되면 갑상선암의 성장과 전이가 활성화 될 가능성이 있다. 또한 갑상선암 자체가 췌장암, 폐암 등 암세포에서 Slit 발현이 감소하였던 다른 암종 보다 악성도가 낮은 것이 이러한 차이의 원인일 수도 있다. 한편, 액손 유도 분자중 하나인 netrin1은 DCC (deleted in colorectal cancer) 와 UNC5 transmembrane 수용체와 결합하여 MAPK 및 AKT 경로를 활성화 하여 세포 사멸을 억제하고 세포의 생존을 돕는 것으로 알려져³¹ ^,32) Slit-Robo 경로 활성 증가 자체가 갑상선암 발생 자체에 영향을 주었을 가능성도 배제할 수 없다. 이를 정확히 규명하기 위해서는 Slit-Robo knockdown 및 과발현 모델을 이용한 추가적인 in vitro 및 in vivo 연구가 필요할 것이다. In contrast to normal tissue, the expression of Slit-Robo pathway was significantly increased in cancer tissues than in normal tissues ( ¹⁶ ^, ²⁰ ^{, 25)} . In this study, we could not find the cause of the increased expression of Slit2 in cancer tissues compared to normal. However, Slit administration showed a clear tumor growth and metastasis inhibitory effect in the thyroid cancer cell line model, and Slit-Robo pathway activity in the thyroid tissue cohort Of the patients with thyroid cancer were found to be associated with a poor prognostic factor, an increase in Slit-Robo pathway activity in thyroid cancer may be the result of not developing thyroid cancer. In other words, the mutation of the other unknown pathway causes the development of thyroid cancer to be secondary to inhibit the Slit-Robo pathway. If this increase is suppressed, the growth and metastasis of thyroid cancer may be activated. In addition, the difference in malignancy between thyroid carcinoma and pancreatic cancer, lung cancer, and other carcinomas may be the cause of this difference. It is known that netrin1, which is one of the axons-inducing molecules, binds to deleted in colorectal cancer (DCC) and UNC5 transmembrane receptors to activate MAPK and AKT pathways, thereby inhibiting apoptosis and helping cell survival ³¹ ^{, 32)} Slit-Robo It is also possible that pathway activity itself may have affected the outbreak of thyroid cancer itself. Additional in vitro and in vivo studies using the Slit-Robo knockdown and overexpression model will be needed to pinpoint this.

서론에서 밝힌 바와 같이 종양 형성과 암세포 이동능에 대한 Slit-Robo 경로의 역할은 단순하지가 않아 각 연구마다 상이한 결과를 보여 주고 있다. 이에 Slit-Robo 경로는 각 암종에 따라, 같은 암종이라 할지라도 그 단계에 따라 각기 다른 영향을 미치고, 각각 Slit과 Robo 아형의 상호작용에 따라서도 각각 다른 역할을 하는 것으로 생각된다¹⁵ ⁾. 때문에 다른 암종의 연구 결과를 갑상선암에 그대로 적용하기에는 어려움이 있다. 본 연구는 처음으로 Slit-Robo 경로의 역할을 다각적으로 분석하였고 갑상선암에서 Slit2가 암세포 이동 억제 능력을 가진 종양 억제 유전자로서의 역할이 있음을 밝힌 최초의 연구라는 점에서 그 의의가 크다.
As noted in the introduction, the role of the Slit-Robo pathway in tumorigenesis and cancer cell migration is not simple, and results in different studies. The Slit-Robo pathway is thought to play different roles, respectively, depending on tumor as also having a different effect according to the stage, each of Slit and Robo interaction subtypes matter such, for each carcinoma ^{^15).} Therefore, it is difficult to apply the results of other carcinomas to thyroid cancer. In this study, we first analyzed the role of the Slit-Robo pathway in a multifaceted manner, and it is significant in that it is the first study to show that Slit2 plays a role as a tumor suppressor gene with the ability to inhibit tumor cell migration in thyroid cancer.

결론conclusion

Slit-Robo 경로의 활성 저하, 특히 Slit2의 발현 저하는 갑상선암의 림프절 전이, 원격전이와 재발 등의 나쁜 예후와 연관이 있었다. 세포 실험에서Slit의 투여는 갑상선암 세포주의 이동능을 뚜렷이 억제하였으며 이는 beta-catenin 활성의 억제, E-cadherin 활성 유도에 의한 것으로 생각된다. 본 연구는 처음으로 갑상선암에서 Slit-Robo 경로의 활성과 예후의 연관성을 확인한 것으로, 새로운 바이오마커 및 새로운 분자 치료 표적으로 Slit의 가능성을 제안하였다.
The decreased activity of Slit-Robo pathway, especially the decreased expression of Slit2, was associated with poor prognosis such as lymph node metastasis, distant metastasis and recurrence of thyroid cancer. In the cell study, Slit administration markedly inhibited the migration of the thyroid cancer cell line, which is thought to be due to inhibition of beta-catenin activity and induction of E-cadherin activity. This study confirmed the association of prognosis with the activity of Slit-Robo pathway for the first time in thyroid cancer, suggesting the possibility of new biomarkers and slit as a new molecular therapy target.

이상으로 본 발명의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.
While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

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<110> THE ASAN FOUNDATION <120> A composition for preventing or inhibiting a metastasis of a primary thyroid cancer <130> PN140635 <160> 12 <170> KopatentIn 2.0 <210> 1 <211> 1529 <212> PRT <213> slit homolog 2 protein isoform 1 precursor [Homo sapiens] <400> 1 Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val 1 5 10 15 Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys 20 25 30 Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser 35 40 45 Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly 50 55 60 Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His 65 70 75 80 Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg 85 90 95 Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg 100 105 110 Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys 115 120 125 Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg 130 135 140 Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr 145 150 155 160 Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp 165 170 175 Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val 180 185 190 Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser 195 200 205 Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu 210 215 220 Arg Gln Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser 225 230 235 240 His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val 245 250 255 Cys Ser Gly His Gln Ser Phe Met Ala Pro Ser Cys Ser Val Leu His 260 265 270 Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val Asp Cys Arg Gly 275 280 285 Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu Thr Ile Thr Glu 290 295 300 Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro Pro Gly Ala Phe 305 310 315 320 Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser Asn Asn Gln Ile 325 330 335 Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg Ser Leu Asn Ser 340 345 350 Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro Lys Ser Leu Phe 355 360 365 Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn Ala Asn Lys Ile 370 375 380 Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His Asn Leu Asn Leu 385 390 395 400 Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala Lys Gly Thr Phe 405 410 415 Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala Gln Asn Pro Phe 420 425 430 Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr Leu His Thr Asn 435 440 445 Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro Arg Arg Leu Ala 450 455 460 Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe Arg Cys Ser Ala 465 470 475 480 Lys Glu Gln Tyr Phe Ile Pro Gly Thr Glu Asp Tyr Arg Ser Lys Leu 485 490 495 Ser Gly Asp Cys Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys 500 505 510 Glu Gly Thr Thr Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile Pro 515 520 525 Glu His Ile Pro Gln Tyr Thr Ala Glu Leu Arg Leu Asn Asn Asn Glu 530 535 540 Phe Thr Val Leu Glu Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu 545 550 555 560 Arg Lys Ile Asn Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly 565 570 575 Ala Phe Glu Gly Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn 580 585 590 Arg Leu Glu Asn Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu 595 600 605 Lys Thr Leu Met Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp 610 615 620 Ser Phe Ile Gly Leu Ser Ser Val Arg Leu Leu Ser Leu Tyr Asp Asn 625 630 635 640 Gln Ile Thr Thr Val Ala Pro Gly Ala Phe Asp Thr Leu His Ser Leu 645 650 655 Ser Thr Leu Asn Leu Leu Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu 660 665 670 Ala Trp Leu Gly Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn 675 680 685 Pro Arg Cys Gln Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp 690 695 700 Val Ala Ile Gln Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser 705 710 715 720 Cys Ser Pro Leu Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu Asp Thr 725 730 735 Val Val Arg Cys Ser Asn Lys Gly Leu Lys Val Leu Pro Lys Gly Ile 740 745 750 Pro Arg Asp Val Thr Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu 755 760 765 Val Pro Lys Glu Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu 770 775 780 Ser Asn Asn Arg Ile Ser Thr Leu Ser Asn Gln Ser Phe Ser Asn Met 785 790 795 800 Thr Gln Leu Leu Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile 805 810 815 Pro Pro Arg Thr Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu 820 825 830 His Gly Asn Asp Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu 835 840 845 Ser Ala Leu Ser His Leu Ala Ile Gly Ala Asn Pro Leu Tyr Cys Asp 850 855 860 Cys Asn Met Gln Trp Leu Ser Asp Trp Val Lys Ser Glu Tyr Lys Glu 865 870 875 880 Pro Gly Ile Ala Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu 885 890 895 Leu Leu Thr Thr Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp 900 905 910 Val Asn Ile Leu Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys 915 920 925 Asn Asp Gly Thr Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr 930 935 940 Cys Pro Tyr Gly Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala 945 950 955 960 Cys Ile Ser Asn Pro Cys Lys His Gly Gly Thr Cys His Leu Lys Glu 965 970 975 Gly Glu Glu Asp Gly Phe Trp Cys Ile Cys Ala Asp Gly Phe Glu Gly 980 985 990 Glu Asn Cys Glu Val Asn Val Asp Asp Cys Glu Asp Asn Asp Cys Glu 995 1000 1005 Asn Asn Ser Thr Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu Cys 1010 1015 1020 Pro Pro Glu Tyr Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp Phe Cys 1025 1030 1035 1040 Ala Gln Asp Leu Asn Pro Cys Gln His Asp Ser Lys Cys Ile Leu Thr 1045 1050 1055 Pro Lys Gly Phe Lys Cys Asp Cys Thr Pro Gly Tyr Val Gly Glu His 1060 1065 1070 Cys Asp Ile Asp Phe Asp Asp Cys Gln Asp Asn Lys Cys Lys Asn Gly 1075 1080 1085 Ala His Cys Thr Asp Ala Val Asn Gly Tyr Thr Cys Ile Cys Pro Glu 1090 1095 1100 Gly Tyr Ser Gly Leu Phe Cys Glu Phe Ser Pro Pro Met Val Leu Pro 1105 1110 1115 1120 Arg Thr Ser Pro Cys Asp Asn Phe Asp Cys Gln Asn Gly Ala Gln Cys 1125 1130 1135 Ile Val Arg Ile Asn Glu Pro Ile Cys Gln Cys Leu Pro Gly Tyr Gln 1140 1145 1150 Gly Glu Lys Cys Glu Lys Leu Val Ser Val Asn Phe Ile Asn Lys Glu 1155 1160 1165 Ser Tyr Leu Gln Ile Pro Ser Ala Lys Val Arg Pro Gln Thr Asn Ile 1170 1175 1180 Thr Leu Gln Ile Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys 1185 1190 1195 1200 Gly Asp Lys Asp His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Arg 1205 1210 1215 Ala Ser Tyr Asp Thr Gly Ser His Pro Ala Ser Ala Ile Tyr Ser Val 1220 1225 1230 Glu Thr Ile Asn Asp Gly Asn Phe His Ile Val Glu Leu Leu Ala Leu 1235 1240 1245 Asp Gln Ser Leu Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile Ile 1250 1255 1260 Thr Asn Leu Ser Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro Leu Tyr 1265 1270 1275 1280 Val Gly Gly Met Pro Gly Lys Ser Asn Val Ala Ser Leu Arg Gln Ala 1285 1290 1295 Pro Gly Gln Asn Gly Thr Ser Phe His Gly Cys Ile Arg Asn Leu Tyr 1300 1305 1310 Ile Asn Ser Glu Leu Gln Asp Phe Gln Lys Val Pro Met Gln Thr Gly 1315 1320 1325 Ile Leu Pro Gly Cys Glu Pro Cys His Lys Lys Val Cys Ala His Gly 1330 1335 1340 Thr Cys Gln Pro Ser Ser Gln Ala Gly Phe Thr Cys Glu Cys Gln Glu 1345 1350 1355 1360 Gly Trp Met Gly Pro Leu Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu 1365 1370 1375 Gly Asn Lys Cys Val His Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser 1380 1385 1390 Tyr Ser Cys Lys Cys Leu Glu Gly His Gly Gly Val Leu Cys Asp Glu 1395 1400 1405 Glu Glu Asp Leu Phe Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly 1410 1415 1420 Lys Cys Arg Leu Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser 1425 1430 1435 1440 Gly Tyr Thr Gly Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu 1445 1450 1455 Arg Ile Arg Asp Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln 1460 1465 1470 Thr Thr Lys Lys Val Ser Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly 1475 1480 1485 Gly Gln Cys Cys Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe 1490 1495 1500 Glu Cys Thr Asp Gly Ser Ser Phe Val Asp Glu Val Glu Lys Val Val 1505 1510 1515 1520 Lys Cys Gly Cys Thr Arg Cys Val Ser 1525 <210> 2 <211> 1525 <212> PRT <213> slit homolog 2 protein isoform 2 precursor [Homo sapiens] <400> 2 Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val 1 5 10 15 Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys 20 25 30 Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser 35 40 45 Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly 50 55 60 Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His 65 70 75 80 Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg 85 90 95 Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg 100 105 110 Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys 115 120 125 Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg 130 135 140 Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr 145 150 155 160 Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp 165 170 175 Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val 180 185 190 Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser 195 200 205 Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu 210 215 220 Arg Gln Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser 225 230 235 240 His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val 245 250 255 Cys Ser Asp Glu Glu Glu Gly His Gln Ser Phe Met Ala Pro Ser Cys 260 265 270 Ser Val Leu His Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val 275 280 285 Asp Cys Arg Gly Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu 290 295 300 Thr Ile Thr Glu Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro 305 310 315 320 Pro Gly Ala Phe Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser 325 330 335 Asn Asn Gln Ile Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg 340 345 350 Ser Leu Asn Ser Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro 355 360 365 Lys Ser Leu Phe Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn 370 375 380 Ala Asn Lys Ile Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His 385 390 395 400 Asn Leu Asn Leu Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala 405 410 415 Lys Gly Thr Phe Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala 420 425 430 Gln Asn Pro Phe Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr 435 440 445 Leu His Thr Asn Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro 450 455 460 Arg Arg Leu Ala Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe 465 470 475 480 Arg Cys Ser Gly Thr Glu Asp Tyr Arg Ser Lys Leu Ser Gly Asp Cys 485 490 495 Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys Glu Gly Thr Thr 500 505 510 Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile Pro Glu His Ile Pro 515 520 525 Gln Tyr Thr Ala Glu Leu Arg Leu Asn Asn Asn Glu Phe Thr Val Leu 530 535 540 Glu Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu Arg Lys Ile Asn 545 550 555 560 Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly Ala Phe Glu Gly 565 570 575 Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn Arg Leu Glu Asn 580 585 590 Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu Lys Thr Leu Met 595 600 605 Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp Ser Phe Ile Gly 610 615 620 Leu Ser Ser Val Arg Leu Leu Ser Leu Tyr Asp Asn Gln Ile Thr Thr 625 630 635 640 Val Ala Pro Gly Ala Phe Asp Thr Leu His Ser Leu Ser Thr Leu Asn 645 650 655 Leu Leu Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu Ala Trp Leu Gly 660 665 670 Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn Pro Arg Cys Gln 675 680 685 Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp Val Ala Ile Gln 690 695 700 Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser Cys Ser Pro Leu 705 710 715 720 Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu Asp Thr Val Val Arg Cys 725 730 735 Ser Asn Lys Gly Leu Lys Val Leu Pro Lys Gly Ile Pro Arg Asp Val 740 745 750 Thr Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu Val Pro Lys Glu 755 760 765 Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu Ser Asn Asn Arg 770 775 780 Ile Ser Thr Leu Ser Asn Gln Ser Phe Ser Asn Met Thr Gln Leu Leu 785 790 795 800 Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile Pro Pro Arg Thr 805 810 815 Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu His Gly Asn Asp 820 825 830 Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu Ser Ala Leu Ser 835 840 845 His Leu Ala Ile Gly Ala Asn Pro Leu Tyr Cys Asp Cys Asn Met Gln 850 855 860 Trp Leu Ser Asp Trp Val Lys Ser Glu Tyr Lys Glu Pro Gly Ile Ala 865 870 875 880 Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu Leu Leu Thr Thr 885 890 895 Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp Val Asn Ile Leu 900 905 910 Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys Asn Asp Gly Thr 915 920 925 Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr Cys Pro Tyr Gly 930 935 940 Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala Cys Ile Ser Asn 945 950 955 960 Pro Cys Lys His Gly Gly Thr Cys His Leu Lys Glu Gly Glu Glu Asp 965 970 975 Gly Phe Trp Cys Ile Cys Ala Asp Gly Phe Glu Gly Glu Asn Cys Glu 980 985 990 Val Asn Val Asp Asp Cys Glu Asp Asn Asp Cys Glu Asn Asn Ser Thr 995 1000 1005 Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu Cys Pro Pro Glu Tyr 1010 1015 1020 Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp Phe Cys Ala Gln Asp Leu 1025 1030 1035 1040 Asn Pro Cys Gln His Asp Ser Lys Cys Ile Leu Thr Pro Lys Gly Phe 1045 1050 1055 Lys Cys Asp Cys Thr Pro Gly Tyr Val Gly Glu His Cys Asp Ile Asp 1060 1065 1070 Phe Asp Asp Cys Gln Asp Asn Lys Cys Lys Asn Gly Ala His Cys Thr 1075 1080 1085 Asp Ala Val Asn Gly Tyr Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly 1090 1095 1100 Leu Phe Cys Glu Phe Ser Pro Pro Met Val Leu Pro Arg Thr Ser Pro 1105 1110 1115 1120 Cys Asp Asn Phe Asp Cys Gln Asn Gly Ala Gln Cys Ile Val Arg Ile 1125 1130 1135 Asn Glu Pro Ile Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys 1140 1145 1150 Glu Lys Leu Val Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln 1155 1160 1165 Ile Pro Ser Ala Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile 1170 1175 1180 Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys Asp 1185 1190 1195 1200 His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Arg Ala Ser Tyr Asp 1205 1210 1215 Thr Gly Ser His Pro Ala Ser Ala Ile Tyr Ser Val Glu Thr Ile Asn 1220 1225 1230 Asp Gly Asn Phe His Ile Val Glu Leu Leu Ala Leu Asp Gln Ser Leu 1235 1240 1245 Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile Ile Thr Asn Leu Ser 1250 1255 1260 Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro Leu Tyr Val Gly Gly Met 1265 1270 1275 1280 Pro Gly Lys Ser Asn Val Ala Ser Leu Arg Gln Ala Pro Gly Gln Asn 1285 1290 1295 Gly Thr Ser Phe His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu 1300 1305 1310 Leu Gln Asp Phe Gln Lys Val Pro Met Gln Thr Gly Ile Leu Pro Gly 1315 1320 1325 Cys Glu Pro Cys His Lys Lys Val Cys Ala His Gly Thr Cys Gln Pro 1330 1335 1340 Ser Ser Gln Ala Gly Phe Thr Cys Glu Cys Gln Glu Gly Trp Met Gly 1345 1350 1355 1360 Pro Leu Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys 1365 1370 1375 Val His Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys 1380 1385 1390 Cys Leu Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu 1395 1400 1405 Phe Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu 1410 1415 1420 Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr Gly 1425 1430 1435 1440 Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp 1445 1450 1455 Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys 1460 1465 1470 Val Ser Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys 1475 1480 1485 Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp 1490 1495 1500 Gly Ser Ser Phe Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys 1505 1510 1515 1520 Thr Arg Cys Val Ser 1525 <210> 3 <211> 1521 <212> PRT <213> slit homolog 2 protein isoform 3 precursor [Homo sapiens] <400> 3 Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val 1 5 10 15 Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys 20 25 30 Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser 35 40 45 Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly 50 55 60 Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His 65 70 75 80 Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg 85 90 95 Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg 100 105 110 Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys 115 120 125 Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg 130 135 140 Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr 145 150 155 160 Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp 165 170 175 Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val 180 185 190 Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser 195 200 205 Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu 210 215 220 Arg Gln Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser 225 230 235 240 His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val 245 250 255 Cys Ser Gly His Gln Ser Phe Met Ala Pro Ser Cys Ser Val Leu His 260 265 270 Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val Asp Cys Arg Gly 275 280 285 Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu Thr Ile Thr Glu 290 295 300 Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro Pro Gly Ala Phe 305 310 315 320 Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser Asn Asn Gln Ile 325 330 335 Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg Ser Leu Asn Ser 340 345 350 Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro Lys Ser Leu Phe 355 360 365 Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn Ala Asn Lys Ile 370 375 380 Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His Asn Leu Asn Leu 385 390 395 400 Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala Lys Gly Thr Phe 405 410 415 Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala Gln Asn Pro Phe 420 425 430 Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr Leu His Thr Asn 435 440 445 Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro Arg Arg Leu Ala 450 455 460 Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe Arg Cys Ser Gly 465 470 475 480 Thr Glu Asp Tyr Arg Ser Lys Leu Ser Gly Asp Cys Phe Ala Asp Leu 485 490 495 Ala Cys Pro Glu Lys Cys Arg Cys Glu Gly Thr Thr Val Asp Cys Ser 500 505 510 Asn Gln Lys Leu Asn Lys Ile Pro Glu His Ile Pro Gln Tyr Thr Ala 515 520 525 Glu Leu Arg Leu Asn Asn Asn Glu Phe Thr Val Leu Glu Ala Thr Gly 530 535 540 Ile Phe Lys Lys Leu Pro Gln Leu Arg Lys Ile Asn Phe Ser Asn Asn 545 550 555 560 Lys Ile Thr Asp Ile Glu Glu Gly Ala Phe Glu Gly Ala Ser Gly Val 565 570 575 Asn Glu Ile Leu Leu Thr Ser Asn Arg Leu Glu Asn Val Gln His Lys 580 585 590 Met Phe Lys Gly Leu Glu Ser Leu Lys Thr Leu Met Leu Arg Ser Asn 595 600 605 Arg Ile Thr Cys Val Gly Asn Asp Ser Phe Ile Gly Leu Ser Ser Val 610 615 620 Arg Leu Leu Ser Leu Tyr Asp Asn Gln Ile Thr Thr Val Ala Pro Gly 625 630 635 640 Ala Phe Asp Thr Leu His Ser Leu Ser Thr Leu Asn Leu Leu Ala Asn 645 650 655 Pro Phe Asn Cys Asn Cys Tyr Leu Ala Trp Leu Gly Glu Trp Leu Arg 660 665 670 Lys Lys Arg Ile Val Thr Gly Asn Pro Arg Cys Gln Lys Pro Tyr Phe 675 680 685 Leu Lys Glu Ile Pro Ile Gln Asp Val Ala Ile Gln Asp Phe Thr Cys 690 695 700 Asp Asp Gly Asn Asp Asp Asn Ser Cys Ser Pro Leu Ser Arg Cys Pro 705 710 715 720 Thr Glu Cys Thr Cys Leu Asp Thr Val Val Arg Cys Ser Asn Lys Gly 725 730 735 Leu Lys Val Leu Pro Lys Gly Ile Pro Arg Asp Val Thr Glu Leu Tyr 740 745 750 Leu Asp Gly Asn Gln Phe Thr Leu Val Pro Lys Glu Leu Ser Asn Tyr 755 760 765 Lys His Leu Thr Leu Ile Asp Leu Ser Asn Asn Arg Ile Ser Thr Leu 770 775 780 Ser Asn Gln Ser Phe Ser Asn Met Thr Gln Leu Leu Thr Leu Ile Leu 785 790 795 800 Ser Tyr Asn Arg Leu Arg Cys Ile Pro Pro Arg Thr Phe Asp Gly Leu 805 810 815 Lys Ser Leu Arg Leu Leu Ser Leu His Gly Asn Asp Ile Ser Val Val 820 825 830 Pro Glu Gly Ala Phe Asn Asp Leu Ser Ala Leu Ser His Leu Ala Ile 835 840 845 Gly Ala Asn Pro Leu Tyr Cys Asp Cys Asn Met Gln Trp Leu Ser Asp 850 855 860 Trp Val Lys Ser Glu Tyr Lys Glu Pro Gly Ile Ala Arg Cys Ala Gly 865 870 875 880 Pro Gly Glu Met Ala Asp Lys Leu Leu Leu Thr Thr Pro Ser Lys Lys 885 890 895 Phe Thr Cys Gln Gly Pro Val Asp Val Asn Ile Leu Ala Lys Cys Asn 900 905 910 Pro Cys Leu Ser Asn Pro Cys Lys Asn Asp Gly Thr Cys Asn Ser Asp 915 920 925 Pro Val Asp Phe Tyr Arg Cys Thr Cys Pro Tyr Gly Phe Lys Gly Gln 930 935 940 Asp Cys Asp Val Pro Ile His Ala Cys Ile Ser Asn Pro Cys Lys His 945 950 955 960 Gly Gly Thr Cys His Leu Lys Glu Gly Glu Glu Asp Gly Phe Trp Cys 965 970 975 Ile Cys Ala Asp Gly Phe Glu Gly Glu Asn Cys Glu Val Asn Val Asp 980 985 990 Asp Cys Glu Asp Asn Asp Cys Glu Asn Asn Ser Thr Cys Val Asp Gly 995 1000 1005 Ile Asn Asn Tyr Thr Cys Leu Cys Pro Pro Glu Tyr Thr Gly Glu Leu 1010 1015 1020 Cys Glu Glu Lys Leu Asp Phe Cys Ala Gln Asp Leu Asn Pro Cys Gln 1025 1030 1035 1040 His Asp Ser Lys Cys Ile Leu Thr Pro Lys Gly Phe Lys Cys Asp Cys 1045 1050 1055 Thr Pro Gly Tyr Val Gly Glu His Cys Asp Ile Asp Phe Asp Asp Cys 1060 1065 1070 Gln Asp Asn Lys Cys Lys Asn Gly Ala His Cys Thr Asp Ala Val Asn 1075 1080 1085 Gly Tyr Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly Leu Phe Cys Glu 1090 1095 1100 Phe Ser Pro Pro Met Val Leu Pro Arg Thr Ser Pro Cys Asp Asn Phe 1105 1110 1115 1120 Asp Cys Gln Asn Gly Ala Gln Cys Ile Val Arg Ile Asn Glu Pro Ile 1125 1130 1135 Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys Glu Lys Leu Val 1140 1145 1150 Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln Ile Pro Ser Ala 1155 1160 1165 Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile Ala Thr Asp Glu 1170 1175 1180 Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys Asp His Ile Ala Val 1185 1190 1195 1200 Glu Leu Tyr Arg Gly Arg Val Arg Ala Ser Tyr Asp Thr Gly Ser His 1205 1210 1215 Pro Ala Ser Ala Ile Tyr Ser Val Glu Thr Ile Asn Asp Gly Asn Phe 1220 1225 1230 His Ile Val Glu Leu Leu Ala Leu Asp Gln Ser Leu Ser Leu Ser Val 1235 1240 1245 Asp Gly Gly Asn Pro Lys Ile Ile Thr Asn Leu Ser Lys Gln Ser Thr 1250 1255 1260 Leu Asn Phe Asp Ser Pro Leu Tyr Val Gly Gly Met Pro Gly Lys Ser 1265 1270 1275 1280 Asn Val Ala Ser Leu Arg Gln Ala Pro Gly Gln Asn Gly Thr Ser Phe 1285 1290 1295 His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu Leu Gln Asp Phe 1300 1305 1310 Gln Lys Val Pro Met Gln Thr Gly Ile Leu Pro Gly Cys Glu Pro Cys 1315 1320 1325 His Lys Lys Val Cys Ala His Gly Thr Cys Gln Pro Ser Ser Gln Ala 1330 1335 1340 Gly Phe Thr Cys Glu Cys Gln Glu Gly Trp Met Gly Pro Leu Cys Asp 1345 1350 1355 1360 Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys Val His Gly Thr 1365 1370 1375 Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys Cys Leu Glu Gly 1380 1385 1390 His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu Phe Asn Pro Cys 1395 1400 1405 Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu Ser Gly Leu Gly 1410 1415 1420 Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr Gly Asp Ser Cys Asp 1425 1430 1435 1440 Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp Tyr Tyr Gln Lys 1445 1450 1455 Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys Val Ser Arg Leu 1460 1465 1470 Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys Gly Pro Leu Arg 1475 1480 1485 Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp Gly Ser Ser Phe 1490 1495 1500 Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys Thr Arg Cys Val 1505 1510 1515 1520 Ser <210> 4 <211> 6398 <212> DNA <213> Homo sapiens slit homolog 2 (Drosophila) (SLIT2), transcript variant 1, mRNA <400> 4 cggattcatc ctcaggacct aaagttgccc aaggagctcc tgctctgcca gaggagggtg 60 gagagggcgg tgggaggcgt gtgcctgagt gggctctact gccttgttcc atattatttg 120 gtgcacattt tccctggcac tctgggttgc tagccccgcc gggcactggg cctcagacac 180 tgcgcggttc cctcggagca gcaagctaaa gaaagccccc agtgccggcg aggaaggagg 240 cggcggggaa agatgcgcgg cgttggctgg cagatgctgt ccctgtcgct ggggttagtg 300 ctggcgatcc tgaacaaggt ggcaccgcag gcgtgcccgg cgcagtgctc ttgctcgggc 360 agcacagtgg actgtcacgg gctggcgctg cgcagcgtgc ccaggaatat cccccgcaac 420 accgagagac tggatttaaa tggaaataac atcacaagaa ttacgaagac agattttgct 480 ggtcttagac atctaagagt tcttcagctt atggagaata agattagcac cattgaaaga 540 ggagcattcc aggatcttaa agaactagag agactgcgtt taaacagaaa tcaccttcag 600 ctgtttcctg agttgctgtt tcttgggact gcgaagctat acaggcttga tctcagtgaa 660 aaccaaattc aggcaatccc aaggaaagct ttccgtgggg cagttgacat aaaaaatttg 720 caactggatt acaaccagat cagctgtatt gaagatgggg cattcagggc tctccgggac 780 ctggaagtgc tcactctcaa caataacaac attactagac tttctgtggc aagtttcaac 840 catatgccta aacttaggac ttttcgactg cattcaaaca acctgtattg tgactgccac 900 ctggcctggc tctccgactg gcttcgccaa aggcctcggg ttggtctgta cactcagtgt 960 atgggcccct cccacctgag aggccataat gtagccgagg ttcaaaaacg agaatttgtc 1020 tgcagtggtc accagtcatt tatggctcct tcttgtagtg ttttgcactg ccctgccgcc 1080 tgtacctgta gcaacaatat cgtagactgt cgtgggaaag gtctcactga gatccccaca 1140 aatcttccag agaccatcac agaaatacgt ttggaacaga acacaatcaa agtcatccct 1200 cctggagctt tctcaccata taaaaagctt agacgaattg acctgagcaa taatcagatc 1260 tctgaacttg caccagatgc tttccaagga ctacgctctc tgaattcact tgtcctctat 1320 ggaaataaaa tcacagaact ccccaaaagt ttatttgaag gactgttttc cttacagctc 1380 ctattattga atgccaacaa gataaactgc cttcgggtag atgcttttca ggatctccac 1440 aacttgaacc ttctctccct atatgacaac aagcttcaga ccatcgccaa ggggaccttt 1500 tcacctcttc gggccattca aactatgcat ttggcccaga acccctttat ttgtgactgc 1560 catctcaagt ggctagcgga ttatctccat accaacccga ttgagaccag tggtgcccgt 1620 tgcaccagcc cccgccgcct ggcaaacaaa agaattggac agatcaaaag caagaaattc 1680 cgttgttcag ctaaagaaca gtatttcatt ccaggtacag aagattatcg atcaaaatta 1740 agtggagact gctttgcgga tctggcttgc cctgaaaagt gtcgctgtga aggaaccaca 1800 gtagattgct ctaatcaaaa gctcaacaaa atcccggagc acattcccca gtacactgca 1860 gagttgcgtc tcaataataa tgaatttacc gtgttggaag ccacaggaat ctttaagaaa 1920 cttcctcaat tacgtaaaat aaactttagc aacaataaga tcacagatat tgaggaggga 1980 gcatttgaag gagcatctgg tgtaaatgaa atacttctta cgagtaatcg tttggaaaat 2040 gtgcagcata agatgttcaa gggattggaa agcctcaaaa ctttgatgtt gagaagcaat 2100 cgaataacct gtgtggggaa tgacagtttc ataggactca gttctgtgcg tttgctttct 2160 ttgtatgata atcaaattac tacagttgca ccaggggcat ttgatactct ccattcttta 2220 tctactctaa acctcttggc caatcctttt aactgtaact gctacctggc ttggttggga 2280 gagtggctga gaaagaagag aattgtcacg ggaaatccta gatgtcaaaa accatacttc 2340 ctgaaagaaa tacccatcca ggatgtggcc attcaggact tcacttgtga tgacggaaat 2400 gatgacaata gttgctcccc actttctcgc tgtcctactg aatgtacttg cttggataca 2460 gtcgtccgat gtagcaacaa gggtttgaag gtcttgccga aaggtattcc aagagatgtc 2520 acagagttgt atctggatgg aaaccaattt acactggttc ccaaggaact ctccaactac 2580 aaacatttaa cacttataga cttaagtaac aacagaataa gcacgctttc taatcagagc 2640 ttcagcaaca tgacccagct cctcacctta attcttagtt acaaccgtct gagatgtatt 2700 cctcctcgca cctttgatgg attaaagtct cttcgattac tttctctaca tggaaatgac 2760 atttctgttg tgcctgaagg tgctttcaat gatctttctg cattatcaca tctagcaatt 2820 ggagccaacc ctctttactg tgattgtaac atgcagtggt tatccgactg ggtgaagtcg 2880 gaatataagg agcctggaat tgctcgttgt gctggtcctg gagaaatggc agataaactt 2940 ttactcacaa ctccctccaa aaaatttacc tgtcaaggtc ctgtggatgt caatattcta 3000 gctaagtgta acccctgcct atcaaatccg tgtaaaaatg atggcacatg taatagtgat 3060 ccagttgact tttaccgatg cacctgtcca tatggtttca aggggcagga ctgtgatgtc 3120 ccaattcatg cctgcatcag taacccatgt aaacatggag gaacttgcca cttaaaggaa 3180 ggagaagaag atggattctg gtgtatttgt gctgatggat ttgaaggaga aaattgtgaa 3240 gtcaacgttg atgattgtga agataatgac tgtgaaaata attctacatg tgtcgatggc 3300 attaataact acacatgcct ttgcccacct gagtatacag gtgagttgtg tgaggagaag 3360 ctggacttct gtgcccagga cctgaacccc tgccagcacg attcaaagtg catcctaact 3420 ccaaagggat tcaaatgtga ctgcacacca gggtacgtag gtgaacactg cgacatcgat 3480 tttgacgact gccaagacaa caagtgtaaa aacggagccc actgcacaga tgcagtgaac 3540 ggctatacgt gcatatgccc cgaaggttac agtggcttgt tctgtgagtt ttctccaccc 3600 atggtcctcc ctcgtaccag cccctgtgat aattttgatt gtcagaatgg agctcagtgt 3660 atcgtcagaa taaatgagcc aatatgtcag tgtttgcctg gctatcaggg agaaaagtgt 3720 gaaaaattgg ttagtgtgaa ttttataaac aaagagtctt atcttcagat tccttcagcc 3780 aaggttcggc ctcagacgaa cataacactt cagattgcca cagatgaaga cagcggaatc 3840 ctcctgtata agggtgacaa agaccatatc gcggtagaac tctatcgggg gcgtgttcgt 3900 gccagctatg acaccggctc tcatccagct tctgccattt acagtgtgga gacaatcaat 3960 gatggaaact tccacattgt ggaactactt gccttggatc agagtctctc tttgtccgtg 4020 gatggtggga accccaaaat catcactaac ttgtcaaagc agtccactct gaattttgac 4080 tctccactct atgtaggagg catgccaggg aagagtaacg tggcatctct gcgccaggcc 4140 cctgggcaga acggaaccag cttccacggc tgcatccgga acctttacat caacagtgag 4200 ctgcaggact tccagaaggt gccgatgcaa acaggcattt tgcctggctg tgagccatgc 4260 cacaagaagg tgtgtgccca tggcacatgc cagcccagca gccaggcagg cttcacctgc 4320 gagtgccagg aaggatggat ggggcccctc tgtgaccaac ggaccaatga cccttgcctt 4380 ggaaataaat gcgtacatgg cacctgcttg cccatcaatg cgttctccta cagctgtaag 4440 tgcttggagg gccatggagg tgtcctctgt gatgaagagg aggatctgtt taacccatgc 4500 caggcgatca agtgcaagca tgggaagtgc aggctttcag gtctggggca gccctactgt 4560 gaatgcagca gtggatacac gggggacagc tgtgatcgag aaatctcttg tcgaggggaa 4620 aggataagag attattacca aaagcagcag ggctatgctg cttgccaaac aaccaagaag 4680 gtgtcccgat tagagtgcag aggtgggtgt gcaggagggc agtgctgtgg accgctgagg 4740 agcaagcggc ggaaatactc tttcgaatgc actgacggct cctcctttgt ggacgaggtt 4800 gagaaagtgg tgaagtgcgg ctgtacgagg tgtgtgtcct aaacacactc ccggcagctc 4860 tgtctttgga aaaggttgta tacttcttga ccgtgtggga ctaatgaatg cttcatagtg 4920 gaaatatttg aaatatattg taaaatacag aacagactta tttttattat gagaataaag 4980 actttttttc tgcatttgga aaaaaaaaaa aagaaatgct tgaactaaag cttcccctat 5040 gctggagaag tatgaagaaa gatatacctg gagacattag aacagcgatg ggaaccattg 5100 caactcgggt ccatctttgt aacatgctga agacaagcag aagcacatgc acgagggaca 5160 gaggagctac tgtgcactgc tgtgaaattg cccagagcat aaaacctgtg taccctcctt 5220 cacatcaacc aagttcacta ggacatacca agcacatgcg tgtgaatgag gatgcaaggc 5280 aagagaatgg aactccagaa ttcacagata gaacaaatgg atgagaaata tttcatgcaa 5340 aatataaagt gtcctgagga tctgggttcc atttacgaga ggaaatgaaa gtgctaaaat 5400 aaattttatc ttccttttaa atgtcagcat gtcagcagaa gcagcacaca aaagtcttta 5460 ccattttcca gtattaattt ttttgtaata taaatgataa aggagatgat aaagaaccaa 5520 tagattattg ataaataaat tagtaataat atgatttttt gtttctatga gttctaaaca 5580 gccctataca gtattcagtt tccatgagaa atatttattg tatcaaagta cattgtactt 5640 aacattttag gccatccttt tactgttcct tgatgcttta atatatatta atttataatt 5700 atcctgatat ttttgtacat ttttcaaact taaaaatcag gatttttttt tttttttttt 5760 tggcaatagt actaacatag ggttttatct tgggatacat atgtattggt ctgtttgttg 5820 accttgacat ctgatcactg atgtcaatga cctactggcc tcattcagga cacctgcaga 5880 gagtatgcaa agtccgagag aggaaaacag aaatctgtat gtgagatggt cattgtacag 5940 aaagaagtgg cccctctgca acatgtcctc acagaaacga aatggtgtgt agcaatcaac 6000 actagaaagt agaccttttg caaattaata tgtccttgac cttttttgcc cttttgtggg 6060 ggtgaggtgg ggataaaaag actgtcatat caagaactgt gacttttctt tccctcaaac 6120 aataaaactc ctttattatc ttaatgctcc catgttaaca tgtttgctgc taaattacaa 6180 tgtagaattg ataatggttt atagtgaact gtgctcttcc ctcattaaaa tcccagggtg 6240 ccctgtaaag atgcagatgt ttcttcctga aaacttcttt ttttacaaag aaaattagat 6300 gtacatgtat aattcagtgt gctttgtctt tctccagatt aatatcggtt acactgctga 6360 tgtttgtaaa ttaaacagat atttacttca ttaaaaaa 6398 <210> 5 <211> 6386 <212> DNA <213> Homo sapiens slit homolog 2 (Drosophila) (SLIT2), transcript variant 2, mRNA <400> 5 cggattcatc ctcaggacct aaagttgccc aaggagctcc tgctctgcca gaggagggtg 60 gagagggcgg tgggaggcgt gtgcctgagt gggctctact gccttgttcc atattatttg 120 gtgcacattt tccctggcac tctgggttgc tagccccgcc gggcactggg cctcagacac 180 tgcgcggttc cctcggagca gcaagctaaa gaaagccccc agtgccggcg aggaaggagg 240 cggcggggaa agatgcgcgg cgttggctgg cagatgctgt ccctgtcgct ggggttagtg 300 ctggcgatcc tgaacaaggt ggcaccgcag gcgtgcccgg cgcagtgctc ttgctcgggc 360 agcacagtgg actgtcacgg gctggcgctg cgcagcgtgc ccaggaatat cccccgcaac 420 accgagagac tggatttaaa tggaaataac atcacaagaa ttacgaagac agattttgct 480 ggtcttagac atctaagagt tcttcagctt atggagaata agattagcac cattgaaaga 540 ggagcattcc aggatcttaa agaactagag agactgcgtt taaacagaaa tcaccttcag 600 ctgtttcctg agttgctgtt tcttgggact gcgaagctat acaggcttga tctcagtgaa 660 aaccaaattc aggcaatccc aaggaaagct ttccgtgggg cagttgacat aaaaaatttg 720 caactggatt acaaccagat cagctgtatt gaagatgggg cattcagggc tctccgggac 780 ctggaagtgc tcactctcaa caataacaac attactagac tttctgtggc aagtttcaac 840 catatgccta aacttaggac ttttcgactg cattcaaaca acctgtattg tgactgccac 900 ctggcctggc tctccgactg gcttcgccaa aggcctcggg ttggtctgta cactcagtgt 960 atgggcccct cccacctgag aggccataat gtagccgagg ttcaaaaacg agaatttgtc 1020 tgcagtgatg aggaagaagg tcaccagtca tttatggctc cttcttgtag tgttttgcac 1080 tgccctgccg cctgtacctg tagcaacaat atcgtagact gtcgtgggaa aggtctcact 1140 gagatcccca caaatcttcc agagaccatc acagaaatac gtttggaaca gaacacaatc 1200 aaagtcatcc ctcctggagc tttctcacca tataaaaagc ttagacgaat tgacctgagc 1260 aataatcaga tctctgaact tgcaccagat gctttccaag gactacgctc tctgaattca 1320 cttgtcctct atggaaataa aatcacagaa ctccccaaaa gtttatttga aggactgttt 1380 tccttacagc tcctattatt gaatgccaac aagataaact gccttcgggt agatgctttt 1440 caggatctcc acaacttgaa ccttctctcc ctatatgaca acaagcttca gaccatcgcc 1500 aaggggacct tttcacctct tcgggccatt caaactatgc atttggccca gaaccccttt 1560 atttgtgact gccatctcaa gtggctagcg gattatctcc ataccaaccc gattgagacc 1620 agtggtgccc gttgcaccag cccccgccgc ctggcaaaca aaagaattgg acagatcaaa 1680 agcaagaaat tccgttgttc aggtacagaa gattatcgat caaaattaag tggagactgc 1740 tttgcggatc tggcttgccc tgaaaagtgt cgctgtgaag gaaccacagt agattgctct 1800 aatcaaaagc tcaacaaaat cccggagcac attccccagt acactgcaga gttgcgtctc 1860 aataataatg aatttaccgt gttggaagcc acaggaatct ttaagaaact tcctcaatta 1920 cgtaaaataa actttagcaa caataagatc acagatattg aggagggagc atttgaagga 1980 gcatctggtg taaatgaaat acttcttacg agtaatcgtt tggaaaatgt gcagcataag 2040 atgttcaagg gattggaaag cctcaaaact ttgatgttga gaagcaatcg aataacctgt 2100 gtggggaatg acagtttcat aggactcagt tctgtgcgtt tgctttcttt gtatgataat 2160 caaattacta cagttgcacc aggggcattt gatactctcc attctttatc tactctaaac 2220 ctcttggcca atccttttaa ctgtaactgc tacctggctt ggttgggaga gtggctgaga 2280 aagaagagaa ttgtcacggg aaatcctaga tgtcaaaaac catacttcct gaaagaaata 2340 cccatccagg atgtggccat tcaggacttc acttgtgatg acggaaatga tgacaatagt 2400 tgctccccac tttctcgctg tcctactgaa tgtacttgct tggatacagt cgtccgatgt 2460 agcaacaagg gtttgaaggt cttgccgaaa ggtattccaa gagatgtcac agagttgtat 2520 ctggatggaa accaatttac actggttccc aaggaactct ccaactacaa acatttaaca 2580 cttatagact taagtaacaa cagaataagc acgctttcta atcagagctt cagcaacatg 2640 acccagctcc tcaccttaat tcttagttac aaccgtctga gatgtattcc tcctcgcacc 2700 tttgatggat taaagtctct tcgattactt tctctacatg gaaatgacat ttctgttgtg 2760 cctgaaggtg ctttcaatga tctttctgca ttatcacatc tagcaattgg agccaaccct 2820 ctttactgtg attgtaacat gcagtggtta tccgactggg tgaagtcgga atataaggag 2880 cctggaattg ctcgttgtgc tggtcctgga gaaatggcag ataaactttt actcacaact 2940 ccctccaaaa aatttacctg tcaaggtcct gtggatgtca atattctagc taagtgtaac 3000 ccctgcctat caaatccgtg taaaaatgat ggcacatgta atagtgatcc agttgacttt 3060 taccgatgca cctgtccata tggtttcaag gggcaggact gtgatgtccc aattcatgcc 3120 tgcatcagta acccatgtaa acatggagga acttgccact taaaggaagg agaagaagat 3180 ggattctggt gtatttgtgc tgatggattt gaaggagaaa attgtgaagt caacgttgat 3240 gattgtgaag ataatgactg tgaaaataat tctacatgtg tcgatggcat taataactac 3300 acatgccttt gcccacctga gtatacaggt gagttgtgtg aggagaagct ggacttctgt 3360 gcccaggacc tgaacccctg ccagcacgat tcaaagtgca tcctaactcc aaagggattc 3420 aaatgtgact gcacaccagg gtacgtaggt gaacactgcg acatcgattt tgacgactgc 3480 caagacaaca agtgtaaaaa cggagcccac tgcacagatg cagtgaacgg ctatacgtgc 3540 atatgccccg aaggttacag tggcttgttc tgtgagtttt ctccacccat ggtcctccct 3600 cgtaccagcc cctgtgataa ttttgattgt cagaatggag ctcagtgtat cgtcagaata 3660 aatgagccaa tatgtcagtg tttgcctggc tatcagggag aaaagtgtga aaaattggtt 3720 agtgtgaatt ttataaacaa agagtcttat cttcagattc cttcagccaa ggttcggcct 3780 cagacgaaca taacacttca gattgccaca gatgaagaca gcggaatcct cctgtataag 3840 ggtgacaaag accatatcgc ggtagaactc tatcgggggc gtgttcgtgc cagctatgac 3900 accggctctc atccagcttc tgccatttac agtgtggaga caatcaatga tggaaacttc 3960 cacattgtgg aactacttgc cttggatcag agtctctctt tgtccgtgga tggtgggaac 4020 cccaaaatca tcactaactt gtcaaagcag tccactctga attttgactc tccactctat 4080 gtaggaggca tgccagggaa gagtaacgtg gcatctctgc gccaggcccc tgggcagaac 4140 ggaaccagct tccacggctg catccggaac ctttacatca acagtgagct gcaggacttc 4200 cagaaggtgc cgatgcaaac aggcattttg cctggctgtg agccatgcca caagaaggtg 4260 tgtgcccatg gcacatgcca gcccagcagc caggcaggct tcacctgcga gtgccaggaa 4320 ggatggatgg ggcccctctg tgaccaacgg accaatgacc cttgccttgg aaataaatgc 4380 gtacatggca cctgcttgcc catcaatgcg ttctcctaca gctgtaagtg cttggagggc 4440 catggaggtg tcctctgtga tgaagaggag gatctgttta acccatgcca ggcgatcaag 4500 tgcaagcatg ggaagtgcag gctttcaggt ctggggcagc cctactgtga atgcagcagt 4560 ggatacacgg gggacagctg tgatcgagaa atctcttgtc gaggggaaag gataagagat 4620 tattaccaaa agcagcaggg ctatgctgct tgccaaacaa ccaagaaggt gtcccgatta 4680 gagtgcagag gtgggtgtgc aggagggcag tgctgtggac cgctgaggag caagcggcgg 4740 aaatactctt tcgaatgcac tgacggctcc tcctttgtgg acgaggttga gaaagtggtg 4800 aagtgcggct gtacgaggtg tgtgtcctaa acacactccc ggcagctctg tctttggaaa 4860 aggttgtata cttcttgacc gtgtgggact aatgaatgct tcatagtgga aatatttgaa 4920 atatattgta aaatacagaa cagacttatt tttattatga gaataaagac tttttttctg 4980 catttggaaa aaaaaaaaaa gaaatgcttg aactaaagct tcccctatgc tggagaagta 5040 tgaagaaaga tatacctgga gacattagaa cagcgatggg aaccattgca actcgggtcc 5100 atctttgtaa catgctgaag acaagcagaa gcacatgcac gagggacaga ggagctactg 5160 tgcactgctg tgaaattgcc cagagcataa aacctgtgta ccctccttca catcaaccaa 5220 gttcactagg acataccaag cacatgcgtg tgaatgagga tgcaaggcaa gagaatggaa 5280 ctccagaatt cacagataga acaaatggat gagaaatatt tcatgcaaaa tataaagtgt 5340 cctgaggatc tgggttccat ttacgagagg aaatgaaagt gctaaaataa attttatctt 5400 ccttttaaat gtcagcatgt cagcagaagc agcacacaaa agtctttacc attttccagt 5460 attaattttt ttgtaatata aatgataaag gagatgataa agaaccaata gattattgat 5520 aaataaatta gtaataatat gattttttgt ttctatgagt tctaaacagc cctatacagt 5580 attcagtttc catgagaaat atttattgta tcaaagtaca ttgtacttaa cattttaggc 5640 catcctttta ctgttccttg atgctttaat atatattaat ttataattat cctgatattt 5700 ttgtacattt ttcaaactta aaaatcagga tttttttttt tttttttttg gcaatagtac 5760 taacataggg ttttatcttg ggatacatat gtattggtct gtttgttgac cttgacatct 5820 gatcactgat gtcaatgacc tactggcctc attcaggaca cctgcagaga gtatgcaaag 5880 tccgagagag gaaaacagaa atctgtatgt gagatggtca ttgtacagaa agaagtggcc 5940 cctctgcaac atgtcctcac agaaacgaaa tggtgtgtag caatcaacac tagaaagtag 6000 accttttgca aattaatatg tccttgacct tttttgccct tttgtggggg tgaggtgggg 6060 ataaaaagac tgtcatatca agaactgtga cttttctttc cctcaaacaa taaaactcct 6120 ttattatctt aatgctccca tgttaacatg tttgctgcta aattacaatg tagaattgat 6180 aatggtttat agtgaactgt gctcttccct cattaaaatc ccagggtgcc ctgtaaagat 6240 gcagatgttt cttcctgaaa acttcttttt ttacaaagaa aattagatgt acatgtataa 6300 ttcagtgtgc tttgtctttc tccagattaa tatcggttac actgctgatg tttgtaaatt 6360 aaacagatat ttacttcatt aaaaaa 6386 <210> 6 <211> 6374 <212> DNA <213> Homo sapiens slit homolog 2 (Drosophila) (SLIT2), transcript variant 3, mRNA <400> 6 cggattcatc ctcaggacct aaagttgccc aaggagctcc tgctctgcca gaggagggtg 60 gagagggcgg tgggaggcgt gtgcctgagt gggctctact gccttgttcc atattatttg 120 gtgcacattt tccctggcac tctgggttgc tagccccgcc gggcactggg cctcagacac 180 tgcgcggttc cctcggagca gcaagctaaa gaaagccccc agtgccggcg aggaaggagg 240 cggcggggaa agatgcgcgg cgttggctgg cagatgctgt ccctgtcgct ggggttagtg 300 ctggcgatcc tgaacaaggt ggcaccgcag gcgtgcccgg cgcagtgctc ttgctcgggc 360 agcacagtgg actgtcacgg gctggcgctg cgcagcgtgc ccaggaatat cccccgcaac 420 accgagagac tggatttaaa tggaaataac atcacaagaa ttacgaagac agattttgct 480 ggtcttagac atctaagagt tcttcagctt atggagaata agattagcac cattgaaaga 540 ggagcattcc aggatcttaa agaactagag agactgcgtt taaacagaaa tcaccttcag 600 ctgtttcctg agttgctgtt tcttgggact gcgaagctat acaggcttga tctcagtgaa 660 aaccaaattc aggcaatccc aaggaaagct ttccgtgggg cagttgacat aaaaaatttg 720 caactggatt acaaccagat cagctgtatt gaagatgggg cattcagggc tctccgggac 780 ctggaagtgc tcactctcaa caataacaac attactagac tttctgtggc aagtttcaac 840 catatgccta aacttaggac ttttcgactg cattcaaaca acctgtattg tgactgccac 900 ctggcctggc tctccgactg gcttcgccaa aggcctcggg ttggtctgta cactcagtgt 960 atgggcccct cccacctgag aggccataat gtagccgagg ttcaaaaacg agaatttgtc 1020 tgcagtggtc accagtcatt tatggctcct tcttgtagtg ttttgcactg ccctgccgcc 1080 tgtacctgta gcaacaatat cgtagactgt cgtgggaaag gtctcactga gatccccaca 1140 aatcttccag agaccatcac agaaatacgt ttggaacaga acacaatcaa agtcatccct 1200 cctggagctt tctcaccata taaaaagctt agacgaattg acctgagcaa taatcagatc 1260 tctgaacttg caccagatgc tttccaagga ctacgctctc tgaattcact tgtcctctat 1320 ggaaataaaa tcacagaact ccccaaaagt ttatttgaag gactgttttc cttacagctc 1380 ctattattga atgccaacaa gataaactgc cttcgggtag atgcttttca ggatctccac 1440 aacttgaacc ttctctccct atatgacaac aagcttcaga ccatcgccaa ggggaccttt 1500 tcacctcttc gggccattca aactatgcat ttggcccaga acccctttat ttgtgactgc 1560 catctcaagt ggctagcgga ttatctccat accaacccga ttgagaccag tggtgcccgt 1620 tgcaccagcc cccgccgcct ggcaaacaaa agaattggac agatcaaaag caagaaattc 1680 cgttgttcag gtacagaaga ttatcgatca aaattaagtg gagactgctt tgcggatctg 1740 gcttgccctg aaaagtgtcg ctgtgaagga accacagtag attgctctaa tcaaaagctc 1800 aacaaaatcc cggagcacat tccccagtac actgcagagt tgcgtctcaa taataatgaa 1860 tttaccgtgt tggaagccac aggaatcttt aagaaacttc ctcaattacg taaaataaac 1920 tttagcaaca ataagatcac agatattgag gagggagcat ttgaaggagc atctggtgta 1980 aatgaaatac ttcttacgag taatcgtttg gaaaatgtgc agcataagat gttcaaggga 2040 ttggaaagcc tcaaaacttt gatgttgaga agcaatcgaa taacctgtgt ggggaatgac 2100 agtttcatag gactcagttc tgtgcgtttg ctttctttgt atgataatca aattactaca 2160 gttgcaccag gggcatttga tactctccat tctttatcta ctctaaacct cttggccaat 2220 ccttttaact gtaactgcta cctggcttgg ttgggagagt ggctgagaaa gaagagaatt 2280 gtcacgggaa atcctagatg tcaaaaacca tacttcctga aagaaatacc catccaggat 2340 gtggccattc aggacttcac ttgtgatgac ggaaatgatg acaatagttg ctccccactt 2400 tctcgctgtc ctactgaatg tacttgcttg gatacagtcg tccgatgtag caacaagggt 2460 ttgaaggtct tgccgaaagg tattccaaga gatgtcacag agttgtatct ggatggaaac 2520 caatttacac tggttcccaa ggaactctcc aactacaaac atttaacact tatagactta 2580 agtaacaaca gaataagcac gctttctaat cagagcttca gcaacatgac ccagctcctc 2640 accttaattc ttagttacaa ccgtctgaga tgtattcctc ctcgcacctt tgatggatta 2700 aagtctcttc gattactttc tctacatgga aatgacattt ctgttgtgcc tgaaggtgct 2760 ttcaatgatc tttctgcatt atcacatcta gcaattggag ccaaccctct ttactgtgat 2820 tgtaacatgc agtggttatc cgactgggtg aagtcggaat ataaggagcc tggaattgct 2880 cgttgtgctg gtcctggaga aatggcagat aaacttttac tcacaactcc ctccaaaaaa 2940 tttacctgtc aaggtcctgt ggatgtcaat attctagcta agtgtaaccc ctgcctatca 3000 aatccgtgta aaaatgatgg cacatgtaat agtgatccag ttgactttta ccgatgcacc 3060 tgtccatatg gtttcaaggg gcaggactgt gatgtcccaa ttcatgcctg catcagtaac 3120 ccatgtaaac atggaggaac ttgccactta aaggaaggag aagaagatgg attctggtgt 3180 atttgtgctg atggatttga aggagaaaat tgtgaagtca acgttgatga ttgtgaagat 3240 aatgactgtg aaaataattc tacatgtgtc gatggcatta ataactacac atgcctttgc 3300 ccacctgagt atacaggtga gttgtgtgag gagaagctgg acttctgtgc ccaggacctg 3360 aacccctgcc agcacgattc aaagtgcatc ctaactccaa agggattcaa atgtgactgc 3420 acaccagggt acgtaggtga acactgcgac atcgattttg acgactgcca agacaacaag 3480 tgtaaaaacg gagcccactg cacagatgca gtgaacggct atacgtgcat atgccccgaa 3540 ggttacagtg gcttgttctg tgagttttct ccacccatgg tcctccctcg taccagcccc 3600 tgtgataatt ttgattgtca gaatggagct cagtgtatcg tcagaataaa tgagccaata 3660 tgtcagtgtt tgcctggcta tcagggagaa aagtgtgaaa aattggttag tgtgaatttt 3720 ataaacaaag agtcttatct tcagattcct tcagccaagg ttcggcctca gacgaacata 3780 acacttcaga ttgccacaga tgaagacagc ggaatcctcc tgtataaggg tgacaaagac 3840 catatcgcgg tagaactcta tcgggggcgt gttcgtgcca gctatgacac cggctctcat 3900 ccagcttctg ccatttacag tgtggagaca atcaatgatg gaaacttcca cattgtggaa 3960 ctacttgcct tggatcagag tctctctttg tccgtggatg gtgggaaccc caaaatcatc 4020 actaacttgt caaagcagtc cactctgaat tttgactctc cactctatgt aggaggcatg 4080 ccagggaaga gtaacgtggc atctctgcgc caggcccctg ggcagaacgg aaccagcttc 4140 cacggctgca tccggaacct ttacatcaac agtgagctgc aggacttcca gaaggtgccg 4200 atgcaaacag gcattttgcc tggctgtgag ccatgccaca agaaggtgtg tgcccatggc 4260 acatgccagc ccagcagcca ggcaggcttc acctgcgagt gccaggaagg atggatgggg 4320 cccctctgtg accaacggac caatgaccct tgccttggaa ataaatgcgt acatggcacc 4380 tgcttgccca tcaatgcgtt ctcctacagc tgtaagtgct tggagggcca tggaggtgtc 4440 ctctgtgatg aagaggagga tctgtttaac ccatgccagg cgatcaagtg caagcatggg 4500 aagtgcaggc tttcaggtct ggggcagccc tactgtgaat gcagcagtgg atacacgggg 4560 gacagctgtg atcgagaaat ctcttgtcga ggggaaagga taagagatta ttaccaaaag 4620 cagcagggct atgctgcttg ccaaacaacc aagaaggtgt cccgattaga gtgcagaggt 4680 gggtgtgcag gagggcagtg ctgtggaccg ctgaggagca agcggcggaa atactctttc 4740 gaatgcactg acggctcctc ctttgtggac gaggttgaga aagtggtgaa gtgcggctgt 4800 acgaggtgtg tgtcctaaac acactcccgg cagctctgtc tttggaaaag gttgtatact 4860 tcttgaccgt gtgggactaa tgaatgcttc atagtggaaa tatttgaaat atattgtaaa 4920 atacagaaca gacttatttt tattatgaga ataaagactt tttttctgca tttggaaaaa 4980 aaaaaaaaga aatgcttgaa ctaaagcttc ccctatgctg gagaagtatg aagaaagata 5040 tacctggaga cattagaaca gcgatgggaa ccattgcaac tcgggtccat ctttgtaaca 5100 tgctgaagac aagcagaagc acatgcacga gggacagagg agctactgtg cactgctgtg 5160 aaattgccca gagcataaaa cctgtgtacc ctccttcaca tcaaccaagt tcactaggac 5220 ataccaagca catgcgtgtg aatgaggatg caaggcaaga gaatggaact ccagaattca 5280 cagatagaac aaatggatga gaaatatttc atgcaaaata taaagtgtcc tgaggatctg 5340 ggttccattt acgagaggaa atgaaagtgc taaaataaat tttatcttcc ttttaaatgt 5400 cagcatgtca gcagaagcag cacacaaaag tctttaccat tttccagtat taattttttt 5460 gtaatataaa tgataaagga gatgataaag aaccaataga ttattgataa ataaattagt 5520 aataatatga ttttttgttt ctatgagttc taaacagccc tatacagtat tcagtttcca 5580 tgagaaatat ttattgtatc aaagtacatt gtacttaaca ttttaggcca tccttttact 5640 gttccttgat gctttaatat atattaattt ataattatcc tgatattttt gtacattttt 5700 caaacttaaa aatcaggatt tttttttttt tttttttggc aatagtacta acatagggtt 5760 ttatcttggg atacatatgt attggtctgt ttgttgacct tgacatctga tcactgatgt 5820 caatgaccta ctggcctcat tcaggacacc tgcagagagt atgcaaagtc cgagagagga 5880 aaacagaaat ctgtatgtga gatggtcatt gtacagaaag aagtggcccc tctgcaacat 5940 gtcctcacag aaacgaaatg gtgtgtagca atcaacacta gaaagtagac cttttgcaaa 6000 ttaatatgtc cttgaccttt tttgcccttt tgtgggggtg aggtggggat aaaaagactg 6060 tcatatcaag aactgtgact tttctttccc tcaaacaata aaactccttt attatcttaa 6120 tgctcccatg ttaacatgtt tgctgctaaa ttacaatgta gaattgataa tggtttatag 6180 tgaactgtgc tcttccctca ttaaaatccc agggtgccct gtaaagatgc agatgtttct 6240 tcctgaaaac ttcttttttt acaaagaaaa ttagatgtac atgtataatt cagtgtgctt 6300 tgtctttctc cagattaata tcggttacac tgctgatgtt tgtaaattaa acagatattt 6360 acttcattaa aaaa 6374 <210> 7 <211> 1651 <212> PRT <213> roundabout homolog 1 isoform a precursor [Homo sapiens] <400> 7 Met Lys Trp Lys His Val Pro Phe Leu Val Met Ile Ser Leu Leu Ser 1 5 10 15 Leu Ser Pro Asn His Leu Phe Leu Ala Gln Leu Ile Pro Asp Pro Glu 20 25 30 Asp Val Glu Arg Gly Asn Asp His Gly Thr Pro Ile Pro Thr Ser Asp 35 40 45 Asn Asp Asp Asn Ser Leu Gly Tyr Thr Gly Ser Arg Leu Arg Gln Glu 50 55 60 Asp Phe Pro Pro Arg Ile Val Glu His Pro Ser Asp Leu Ile Val Ser 65 70 75 80 Lys Gly Glu Pro Ala Thr Leu Asn Cys Lys Ala Glu Gly Arg Pro Thr 85 90 95 Pro Thr Ile Glu Trp Tyr Lys Gly Gly Glu Arg Val Glu Thr Asp Lys 100 105 110 Asp Asp Pro Arg Ser His Arg Met Leu Leu Pro Ser Gly Ser Leu Phe 115 120 125 Phe Leu Arg Ile Val His Gly Arg Lys Ser Arg Pro Asp Glu Gly Val 130 135 140 Tyr Val Cys Val Ala Arg Asn Tyr Leu Gly Glu Ala Val Ser His Asn 145 150 155 160 Ala Ser Leu Glu Val Ala Ile Leu Arg Asp Asp Phe Arg Gln Asn Pro 165 170 175 Ser Asp Val Met Val Ala Val Gly Glu Pro Ala Val Met Glu Cys Gln 180 185 190 Pro Pro Arg Gly His Pro Glu Pro Thr Ile Ser Trp Lys Lys Asp Gly 195 200 205 Ser Pro Leu Asp Asp Lys Asp Glu Arg Ile Thr Ile Arg Gly Gly Lys 210 215 220 Leu Met Ile Thr Tyr Thr Arg Lys Ser Asp Ala Gly Lys Tyr Val Cys 225 230 235 240 Val Gly Thr Asn Met Val Gly Glu Arg Glu Ser Glu Val Ala Glu Leu 245 250 255 Thr Val Leu Glu Arg Pro Ser Phe Val Lys Arg Pro Ser Asn Leu Ala 260 265 270 Val Thr Val Asp Asp Ser Ala Glu Phe Lys Cys Glu Ala Arg Gly Asp 275 280 285 Pro Val Pro Thr Val Arg Trp Arg Lys Asp Asp Gly Glu Leu Pro Lys 290 295 300 Ser Arg Tyr Glu Ile Arg Asp Asp His Thr Leu Lys Ile Arg Lys Val 305 310 315 320 Thr Ala Gly Asp Met Gly Ser Tyr Thr Cys Val Ala Glu Asn Met Val 325 330 335 Gly Lys Ala Glu Ala Ser Ala Thr Leu Thr Val Gln Glu Pro Pro His 340 345 350 Phe Val Val Lys Pro Arg Asp Gln Val Val Ala Leu Gly Arg Thr Val 355 360 365 Thr Phe Gln Cys Glu Ala Thr Gly Asn Pro Gln Pro Ala Ile Phe Trp 370 375 380 Arg Arg Glu Gly Ser Gln Asn Leu Leu Phe Ser Tyr Gln Pro Pro Gln 385 390 395 400 Ser Ser Ser Arg Phe Ser Val Ser Gln Thr Gly Asp Leu Thr Ile Thr 405 410 415 Asn Val Gln Arg Ser Asp Val Gly Tyr Tyr Ile Cys Gln Thr Leu Asn 420 425 430 Val Ala Gly Ser Ile Ile Thr Lys Ala Tyr Leu Glu Val Thr Asp Val 435 440 445 Ile Ala Asp Arg Pro Pro Pro Val Ile Arg Gln Gly Pro Val Asn Gln 450 455 460 Thr Val Ala Val Asp Gly Thr Phe Val Leu Ser Cys Val Ala Thr Gly 465 470 475 480 Ser Pro Val Pro Thr Ile Leu Trp Arg Lys Asp Gly Val Leu Val Ser 485 490 495 Thr Gln Asp Ser Arg Ile Lys Gln Leu Glu Asn Gly Val Leu Gln Ile 500 505 510 Arg Tyr Ala Lys Leu Gly Asp Thr Gly Arg Tyr Thr Cys Ile Ala Ser 515 520 525 Thr Pro Ser Gly Glu Ala Thr Trp Ser Ala Tyr Ile Glu Val Gln Glu 530 535 540 Phe Gly Val Pro Val Gln Pro Pro Arg Pro Thr Asp Pro Asn Leu Ile 545 550 555 560 Pro Ser Ala Pro Ser Lys Pro Glu Val Thr Asp Val Ser Arg Asn Thr 565 570 575 Val Thr Leu Ser Trp Gln Pro Asn Leu Asn Ser Gly Ala Thr Pro Thr 580 585 590 Ser Tyr Ile Ile Glu Ala Phe Ser His Ala Ser Gly Ser Ser Trp Gln 595 600 605 Thr Val Ala Glu Asn Val Lys Thr Glu Thr Ser Ala Ile Lys Gly Leu 610 615 620 Lys Pro Asn Ala Ile Tyr Leu Phe Leu Val Arg Ala Ala Asn Ala Tyr 625 630 635 640 Gly Ile Ser Asp Pro Ser Gln Ile Ser Asp Pro Val Lys Thr Gln Asp 645 650 655 Val Leu Pro Thr Ser Gln Gly Val Asp His Lys Gln Val Gln Arg Glu 660 665 670 Leu Gly Asn Ala Val Leu His Leu His Asn Pro Thr Val Leu Ser Ser 675 680 685 Ser Ser Ile Glu Val His Trp Thr Val Asp Gln Gln Ser Gln Tyr Ile 690 695 700 Gln Gly Tyr Lys Ile Leu Tyr Arg Pro Ser Gly Ala Asn His Gly Glu 705 710 715 720 Ser Asp Trp Leu Val Phe Glu Val Arg Thr Pro Ala Lys Asn Ser Val 725 730 735 Val Ile Pro Asp Leu Arg Lys Gly Val Asn Tyr Glu Ile Lys Ala Arg 740 745 750 Pro Phe Phe Asn Glu Phe Gln Gly Ala Asp Ser Glu Ile Lys Phe Ala 755 760 765 Lys Thr Leu Glu Glu Ala Pro Ser Ala Pro Pro Gln Gly Val Thr Val 770 775 780 Ser Lys Asn Asp Gly Asn Gly Thr Ala Ile Leu Val Ser Trp Gln Pro 785 790 795 800 Pro Pro Glu Asp Thr Gln Asn Gly Met Val Gln Glu Tyr Lys Val Trp 805 810 815 Cys Leu Gly Asn Glu Thr Arg Tyr His Ile Asn Lys Thr Val Asp Gly 820 825 830 Ser Thr Phe Ser Val Val Ile Pro Phe Leu Val Pro Gly Ile Arg Tyr 835 840 845 Ser Val Glu Val Ala Ala Ser Thr Gly Ala Gly Ser Gly Val Lys Ser 850 855 860 Glu Pro Gln Phe Ile Gln Leu Asp Ala His Gly Asn Pro Val Ser Pro 865 870 875 880 Glu Asp Gln Val Ser Leu Ala Gln Gln Ile Ser Asp Val Val Lys Gln 885 890 895 Pro Ala Phe Ile Ala Gly Ile Gly Ala Ala Cys Trp Ile Ile Leu Met 900 905 910 Val Phe Ser Ile Trp Leu Tyr Arg His Arg Lys Lys Arg Asn Gly Leu 915 920 925 Thr Ser Thr Tyr Ala Gly Ile Arg Lys Val Pro Ser Phe Thr Phe Thr 930 935 940 Pro Thr Val Thr Tyr Gln Arg Gly Gly Glu Ala Val Ser Ser Gly Gly 945 950 955 960 Arg Pro Gly Leu Leu Asn Ile Ser Glu Pro Ala Ala Gln Pro Trp Leu 965 970 975 Ala Asp Thr Trp Pro Asn Thr Gly Asn Asn His Asn Asp Cys Ser Ile 980 985 990 Ser Cys Cys Thr Ala Gly Asn Gly Asn Ser Asp Ser Asn Leu Thr Thr 995 1000 1005 Tyr Ser Arg Pro Ala Asp Cys Ile Ala Asn Tyr Asn Asn Gln Leu Asp 1010 1015 1020 Asn Lys Gln Thr Asn Leu Met Leu Pro Glu Ser Thr Val Tyr Gly Asp 1025 1030 1035 1040 Val Asp Leu Ser Asn Lys Ile Asn Glu Met Lys Thr Phe Asn Ser Pro 1045 1050 1055 Asn Leu Lys Asp Gly Arg Phe Val Asn Pro Ser Gly Gln Pro Thr Pro 1060 1065 1070 Tyr Ala Thr Thr Gln Leu Ile Gln Ser Asn Leu Ser Asn Asn Met Asn 1075 1080 1085 Asn Gly Ser Gly Asp Ser Gly Glu Lys His Trp Lys Pro Leu Gly Gln 1090 1095 1100 Gln Lys Gln Glu Val Ala Pro Val Gln Tyr Asn Ile Val Glu Gln Asn 1105 1110 1115 1120 Lys Leu Asn Lys Asp Tyr Arg Ala Asn Asp Thr Val Pro Pro Thr Ile 1125 1130 1135 Pro Tyr Asn Gln Ser Tyr Asp Gln Asn Thr Gly Gly Ser Tyr Asn Ser 1140 1145 1150 Ser Asp Arg Gly Ser Ser Thr Ser Gly Ser Gln Gly His Lys Lys Gly 1155 1160 1165 Ala Arg Thr Pro Lys Val Pro Lys Gln Gly Gly Met Asn Trp Ala Asp 1170 1175 1180 Leu Leu Pro Pro Pro Pro Ala His Pro Pro Pro His Ser Asn Ser Glu 1185 1190 1195 1200 Glu Tyr Asn Ile Ser Val Asp Glu Ser Tyr Asp Gln Glu Met Pro Cys 1205 1210 1215 Pro Val Pro Pro Ala Arg Met Tyr Leu Gln Gln Asp Glu Leu Glu Glu 1220 1225 1230 Glu Glu Asp Glu Arg Gly Pro Thr Pro Pro Val Arg Gly Ala Ala Ser 1235 1240 1245 Ser Pro Ala Ala Val Ser Tyr Ser His Gln Ser Thr Ala Thr Leu Thr 1250 1255 1260 Pro Ser Pro Gln Glu Glu Leu Gln Pro Met Leu Gln Asp Cys Pro Glu 1265 1270 1275 1280 Glu Thr Gly His Met Gln His Gln Pro Asp Arg Arg Arg Gln Pro Val 1285 1290 1295 Ser Pro Pro Pro Pro Pro Arg Pro Ile Ser Pro Pro His Thr Tyr Gly 1300 1305 1310 Tyr Ile Ser Gly Pro Leu Val Ser Asp Met Asp Thr Asp Ala Pro Glu 1315 1320 1325 Glu Glu Glu Asp Glu Ala Asp Met Glu Val Ala Lys Met Gln Thr Arg 1330 1335 1340 Arg Leu Leu Leu Arg Gly Leu Glu Gln Thr Pro Ala Ser Ser Val Gly 1345 1350 1355 1360 Asp Leu Glu Ser Ser Val Thr Gly Ser Met Ile Asn Gly Trp Gly Ser 1365 1370 1375 Ala Ser Glu Glu Asp Asn Ile Ser Ser Gly Arg Ser Ser Val Ser Ser 1380 1385 1390 Ser Asp Gly Ser Phe Phe Thr Asp Ala Asp Phe Ala Gln Ala Val Ala 1395 1400 1405 Ala Ala Ala Glu Tyr Ala Gly Leu Lys Val Ala Arg Arg Gln Met Gln 1410 1415 1420 Asp Ala Ala Gly Arg Arg His Phe His Ala Ser Gln Cys Pro Arg Pro 1425 1430 1435 1440 Thr Ser Pro Val Ser Thr Asp Ser Asn Met Ser Ala Ala Val Met Gln 1445 1450 1455 Lys Thr Arg Pro Ala Lys Lys Leu Lys His Gln Pro Gly His Leu Arg 1460 1465 1470 Arg Glu Thr Tyr Thr Asp Asp Leu Pro Pro Pro Pro Val Pro Pro Pro 1475 1480 1485 Ala Ile Lys Ser Pro Thr Ala Gln Ser Lys Thr Gln Leu Glu Val Arg 1490 1495 1500 Pro Val Val Val Pro Lys Leu Pro Ser Met Asp Ala Arg Thr Asp Arg 1505 1510 1515 1520 Ser Ser Asp Arg Lys Gly Ser Ser Tyr Lys Gly Arg Glu Val Leu Asp 1525 1530 1535 Gly Arg Gln Val Val Asp Met Arg Thr Asn Pro Gly Asp Pro Arg Glu 1540 1545 1550 Ala Gln Glu Gln Gln Asn Asp Gly Lys Gly Arg Gly Asn Lys Ala Ala 1555 1560 1565 Lys Arg Asp Leu Pro Pro Ala Lys Thr His Leu Ile Gln Glu Asp Ile 1570 1575 1580 Leu Pro Tyr Cys Arg Pro Thr Phe Pro Thr Ser Asn Asn Pro Arg Asp 1585 1590 1595 1600 Pro Ser Ser Ser Ser Ser Met Ser Ser Arg Gly Ser Gly Ser Arg Gln 1605 1610 1615 Arg Glu Gln Ala Asn Val Gly Arg Arg Asn Ile Ala Glu Met Gln Val 1620 1625 1630 Leu Gly Gly Tyr Glu Arg Gly Glu Asp Asn Asn Glu Glu Leu Glu Glu 1635 1640 1645 Thr Glu Ser 1650 <210> 8 <211> 1606 <212> PRT <213> roundabout homolog 1 isoform b [Homo sapiens] <400> 8 Met Ile Ala Glu Pro Ala His Phe Tyr Leu Phe Gly Leu Ile Cys Leu 1 5 10 15 Cys Ser Gly Ser Arg Leu Arg Gln Glu Asp Phe Pro Pro Arg Ile Val 20 25 30 Glu His Pro Ser Asp Leu Ile Val Ser Lys Gly Glu Pro Ala Thr Leu 35 40 45 Asn Cys Lys Ala Glu Gly Arg Pro Thr Pro Thr Ile Glu Trp Tyr Lys 50 55 60 Gly Gly Glu Arg Val Glu Thr Asp Lys Asp Asp Pro Arg Ser His Arg 65 70 75 80 Met Leu Leu Pro Ser Gly Ser Leu Phe Phe Leu Arg Ile Val His Gly 85 90 95 Arg Lys Ser Arg Pro Asp Glu Gly Val Tyr Val Cys Val Ala Arg Asn 100 105 110 Tyr Leu Gly Glu Ala Val Ser His Asn Ala Ser Leu Glu Val Ala Ile 115 120 125 Leu Arg Asp Asp Phe Arg Gln Asn Pro Ser Asp Val Met Val Ala Val 130 135 140 Gly Glu Pro Ala Val Met Glu Cys Gln Pro Pro Arg Gly His Pro Glu 145 150 155 160 Pro Thr Ile Ser Trp Lys Lys Asp Gly Ser Pro Leu Asp Asp Lys Asp 165 170 175 Glu Arg Ile Thr Ile Arg Gly Gly Lys Leu Met Ile Thr Tyr Thr Arg 180 185 190 Lys Ser Asp Ala Gly Lys Tyr Val Cys Val Gly Thr Asn Met Val Gly 195 200 205 Glu Arg Glu Ser Glu Val Ala Glu Leu Thr Val Leu Glu Arg Pro Ser 210 215 220 Phe Val Lys Arg Pro Ser Asn Leu Ala Val Thr Val Asp Asp Ser Ala 225 230 235 240 Glu Phe Lys Cys Glu Ala Arg Gly Asp Pro Val Pro Thr Val Arg Trp 245 250 255 Arg Lys Asp Asp Gly Glu Leu Pro Lys Ser Arg Tyr Glu Ile Arg Asp 260 265 270 Asp His Thr Leu Lys Ile Arg Lys Val Thr Ala Gly Asp Met Gly Ser 275 280 285 Tyr Thr Cys Val Ala Glu Asn Met Val Gly Lys Ala Glu Ala Ser Ala 290 295 300 Thr Leu Thr Val Gln Val Gly Ser Glu Pro Pro His Phe Val Val Lys 305 310 315 320 Pro Arg Asp Gln Val Val Ala Leu Gly Arg Thr Val Thr Phe Gln Cys 325 330 335 Glu Ala Thr Gly Asn Pro Gln Pro Ala Ile Phe Trp Arg Arg Glu Gly 340 345 350 Ser Gln Asn Leu Leu Phe Ser Tyr Gln Pro Pro Gln Ser Ser Ser Arg 355 360 365 Phe Ser Val Ser Gln Thr Gly Asp Leu Thr Ile Thr Asn Val Gln Arg 370 375 380 Ser Asp Val Gly Tyr Tyr Ile Cys Gln Thr Leu Asn Val Ala Gly Ser 385 390 395 400 Ile Ile Thr Lys Ala Tyr Leu Glu Val Thr Asp Val Ile Ala Asp Arg 405 410 415 Pro Pro Pro Val Ile Arg Gln Gly Pro Val Asn Gln Thr Val Ala Val 420 425 430 Asp Gly Thr Phe Val Leu Ser Cys Val Ala Thr Gly Ser Pro Val Pro 435 440 445 Thr Ile Leu Trp Arg Lys Asp Gly Val Leu Val Ser Thr Gln Asp Ser 450 455 460 Arg Ile Lys Gln Leu Glu Asn Gly Val Leu Gln Ile Arg Tyr Ala Lys 465 470 475 480 Leu Gly Asp Thr Gly Arg Tyr Thr Cys Ile Ala Ser Thr Pro Ser Gly 485 490 495 Glu Ala Thr Trp Ser Ala Tyr Ile Glu Val Gln Glu Phe Gly Val Pro 500 505 510 Val Gln Pro Pro Arg Pro Thr Asp Pro Asn Leu Ile Pro Ser Ala Pro 515 520 525 Ser Lys Pro Glu Val Thr Asp Val Ser Arg Asn Thr Val Thr Leu Ser 530 535 540 Trp Gln Pro Asn Leu Asn Ser Gly Ala Thr Pro Thr Ser Tyr Ile Ile 545 550 555 560 Glu Ala Phe Ser His Ala Ser Gly Ser Ser Trp Gln Thr Val Ala Glu 565 570 575 Asn Val Lys Thr Glu Thr Ser Ala Ile Lys Gly Leu Lys Pro Asn Ala 580 585 590 Ile Tyr Leu Phe Leu Val Arg Ala Ala Asn Ala Tyr Gly Ile Ser Asp 595 600 605 Pro Ser Gln Ile Ser Asp Pro Val Lys Thr Gln Asp Val Leu Pro Thr 610 615 620 Ser Gln Gly Val Asp His Lys Gln Val Gln Arg Glu Leu Gly Asn Ala 625 630 635 640 Val Leu His Leu His Asn Pro Thr Val Leu Ser Ser Ser Ser Ile Glu 645 650 655 Val His Trp Thr Val Asp Gln Gln Ser Gln Tyr Ile Gln Gly Tyr Lys 660 665 670 Ile Leu Tyr Arg Pro Ser Gly Ala Asn His Gly Glu Ser Asp Trp Leu 675 680 685 Val Phe Glu Val Arg Thr Pro Ala Lys Asn Ser Val Val Ile Pro Asp 690 695 700 Leu Arg Lys Gly Val Asn Tyr Glu Ile Lys Ala Arg Pro Phe Phe Asn 705 710 715 720 Glu Phe Gln Gly Ala Asp Ser Glu Ile Lys Phe Ala Lys Thr Leu Glu 725 730 735 Glu Ala Pro Ser Ala Pro Pro Gln Gly Val Thr Val Ser Lys Asn Asp 740 745 750 Gly Asn Gly Thr Ala Ile Leu Val Ser Trp Gln Pro Pro Pro Glu Asp 755 760 765 Thr Gln Asn Gly Met Val Gln Glu Tyr Lys Val Trp Cys Leu Gly Asn 770 775 780 Glu Thr Arg Tyr His Ile Asn Lys Thr Val Asp Gly Ser Thr Phe Ser 785 790 795 800 Val Val Ile Pro Phe Leu Val Pro Gly Ile Arg Tyr Ser Val Glu Val 805 810 815 Ala Ala Ser Thr Gly Ala Gly Ser Gly Val Lys Ser Glu Pro Gln Phe 820 825 830 Ile Gln Leu Asp Ala His Gly Asn Pro Val Ser Pro Glu Asp Gln Val 835 840 845 Ser Leu Ala Gln Gln Ile Ser Asp Val Val Lys Gln Pro Ala Phe Ile 850 855 860 Ala Gly Ile Gly Ala Ala Cys Trp Ile Ile Leu Met Val Phe Ser Ile 865 870 875 880 Trp Leu Tyr Arg His Arg Lys Lys Arg Asn Gly Leu Thr Ser Thr Tyr 885 890 895 Ala Gly Ile Arg Lys Val Thr Tyr Gln Arg Gly Gly Glu Ala Val Ser 900 905 910 Ser Gly Gly Arg Pro Gly Leu Leu Asn Ile Ser Glu Pro Ala Ala Gln 915 920 925 Pro Trp Leu Ala Asp Thr Trp Pro Asn Thr Gly Asn Asn His Asn Asp 930 935 940 Cys Ser Ile Ser Cys Cys Thr Ala Gly Asn Gly Asn Ser Asp Ser Asn 945 950 955 960 Leu Thr Thr Tyr Ser Arg Pro Ala Asp Cys Ile Ala Asn Tyr Asn Asn 965 970 975 Gln Leu Asp Asn Lys Gln Thr Asn Leu Met Leu Pro Glu Ser Thr Val 980 985 990 Tyr Gly Asp Val Asp Leu Ser Asn Lys Ile Asn Glu Met Lys Thr Phe 995 1000 1005 Asn Ser Pro Asn Leu Lys Asp Gly Arg Phe Val Asn Pro Ser Gly Gln 1010 1015 1020 Pro Thr Pro Tyr Ala Thr Thr Gln Leu Ile Gln Ser Asn Leu Ser Asn 1025 1030 1035 1040 Asn Met Asn Asn Gly Ser Gly Asp Ser Gly Glu Lys His Trp Lys Pro 1045 1050 1055 Leu Gly Gln Gln Lys Gln Glu Val Ala Pro Val Gln Tyr Asn Ile Val 1060 1065 1070 Glu Gln Asn Lys Leu Asn Lys Asp Tyr Arg Ala Asn Asp Thr Val Pro 1075 1080 1085 Pro Thr Ile Pro Tyr Asn Gln Ser Tyr Asp Gln Asn Thr Gly Gly Ser 1090 1095 1100 Tyr Asn Ser Ser Asp Arg Gly Ser Ser Thr Ser Gly Ser Gln Gly His 1105 1110 1115 1120 Lys Lys Gly Ala Arg Thr Pro Lys Val Pro Lys Gln Gly Gly Met Asn 1125 1130 1135 Trp Ala Asp Leu Leu Pro Pro Pro Pro Ala His Pro Pro Pro His Ser 1140 1145 1150 Asn Ser Glu Glu Tyr Asn Ile Ser Val Asp Glu Ser Tyr Asp Gln Glu 1155 1160 1165 Met Pro Cys Pro Val Pro Pro Ala Arg Met Tyr Leu Gln Gln Asp Glu 1170 1175 1180 Leu Glu Glu Glu Glu Asp Glu Arg Gly Pro Thr Pro Pro Val Arg Gly 1185 1190 1195 1200 Ala Ala Ser Ser Pro Ala Ala Val Ser Tyr Ser His Gln Ser Thr Ala 1205 1210 1215 Thr Leu Thr Pro Ser Pro Gln Glu Glu Leu Gln Pro Met Leu Gln Asp 1220 1225 1230 Cys Pro Glu Glu Thr Gly His Met Gln His Gln Pro Asp Arg Arg Arg 1235 1240 1245 Gln Pro Val Ser Pro Pro Pro Pro Pro Arg Pro Ile Ser Pro Pro His 1250 1255 1260 Thr Tyr Gly Tyr Ile Ser Gly Pro Leu Val Ser Asp Met Asp Thr Asp 1265 1270 1275 1280 Ala Pro Glu Glu Glu Glu Asp Glu Ala Asp Met Glu Val Ala Lys Met 1285 1290 1295 Gln Thr Arg Arg Leu Leu Leu Arg Gly Leu Glu Gln Thr Pro Ala Ser 1300 1305 1310 Ser Val Gly Asp Leu Glu Ser Ser Val Thr Gly Ser Met Ile Asn Gly 1315 1320 1325 Trp Gly Ser Ala Ser Glu Glu Asp Asn Ile Ser Ser Gly Arg Ser Ser 1330 1335 1340 Val Ser Ser Ser Asp Gly Ser Phe Phe Thr Asp Ala Asp Phe Ala Gln 1345 1350 1355 1360 Ala Val Ala Ala Ala Ala Glu Tyr Ala Gly Leu Lys Val Ala Arg Arg 1365 1370 1375 Gln Met Gln Asp Ala Ala Gly Arg Arg His Phe His Ala Ser Gln Cys 1380 1385 1390 Pro Arg Pro Thr Ser Pro Val Ser Thr Asp Ser Asn Met Ser Ala Ala 1395 1400 1405 Val Met Gln Lys Thr Arg Pro Ala Lys Lys Leu Lys His Gln Pro Gly 1410 1415 1420 His Leu Arg Arg Glu Thr Tyr Thr Asp Asp Leu Pro Pro Pro Pro Val 1425 1430 1435 1440 Pro Pro Pro Ala Ile Lys Ser Pro Thr Ala Gln Ser Lys Thr Gln Leu 1445 1450 1455 Glu Val Arg Pro Val Val Val Pro Lys Leu Pro Ser Met Asp Ala Arg 1460 1465 1470 Thr Asp Arg Ser Ser Asp Arg Lys Gly Ser Ser Tyr Lys Gly Arg Glu 1475 1480 1485 Val Leu Asp Gly Arg Gln Val Val Asp Met Arg Thr Asn Pro Gly Asp 1490 1495 1500 Pro Arg Glu Ala Gln Glu Gln Gln Asn Asp Gly Lys Gly Arg Gly Asn 1505 1510 1515 1520 Lys Ala Ala Lys Arg Asp Leu Pro Pro Ala Lys Thr His Leu Ile Gln 1525 1530 1535 Glu Asp Ile Leu Pro Tyr Cys Arg Pro Thr Phe Pro Thr Ser Asn Asn 1540 1545 1550 Pro Arg Asp Pro Ser Ser Ser Ser Ser Met Ser Ser Arg Gly Ser Gly 1555 1560 1565 Ser Arg Gln Arg Glu Gln Ala Asn Val Gly Arg Arg Asn Ile Ala Glu 1570 1575 1580 Met Gln Val Leu Gly Gly Tyr Glu Arg Gly Glu Asp Asn Asn Glu Glu 1585 1590 1595 1600 Leu Glu Glu Thr Glu Ser 1605 <210> 9 <211> 1551 <212> PRT <213> roundabout homolog 1 isoform d [Homo sapiens] <400> 9 Met Ile Ala Glu Pro Ala His Phe Tyr Leu Phe Gly Leu Ile Cys Leu 1 5 10 15 Cys Ser Gly Ser Arg Leu Arg Gln Glu Asp Phe Pro Pro Arg Ile Val 20 25 30 Glu His Pro Ser Asp Leu Ile Val Ser Lys Gly Glu Pro Ala Thr Leu 35 40 45 Asn Cys Lys Ala Glu Gly Arg Pro Thr Pro Thr Ile Glu Trp Tyr Lys 50 55 60 Gly Gly Glu Arg Val Glu Thr Asp Lys Asp Asp Pro Arg Ser His Arg 65 70 75 80 Met Leu Leu Pro Ser Gly Ser Leu Phe Phe Leu Arg Ile Val His Gly 85 90 95 Arg Lys Ser Arg Pro Asp Glu Gly Val Tyr Val Cys Val Ala Arg Asn 100 105 110 Tyr Leu Gly Glu Ala Val Ser His Asn Ala Ser Leu Glu Val Ala Ile 115 120 125 Leu Arg Asp Asp Phe Arg Gln Asn Pro Ser Asp Val Met Val Ala Val 130 135 140 Gly Glu Pro Ala Val Met Glu Cys Gln Pro Pro Arg Gly His Pro Glu 145 150 155 160 Pro Thr Ile Ser Trp Lys Lys Asp Gly Ser Pro Leu Asp Asp Lys Asp 165 170 175 Glu Arg Ile Thr Ile Arg Gly Gly Lys Leu Met Ile Thr Tyr Thr Arg 180 185 190 Lys Ser Asp Ala Gly Lys Tyr Val Cys Val Gly Thr Asn Met Val Gly 195 200 205 Glu Arg Glu Ser Glu Val Ala Glu Leu Thr Val Leu Glu Arg Pro Ser 210 215 220 Phe Val Lys Arg Pro Ser Asn Leu Ala Val Thr Val Asp Asp Ser Ala 225 230 235 240 Glu Phe Lys Cys Glu Ala Arg Gly Asp Pro Val Pro Thr Val Arg Trp 245 250 255 Arg Lys Asp Asp Gly Glu Leu Pro Lys Ser Arg Tyr Glu Ile Arg Asp 260 265 270 Asp His Thr Leu Lys Ile Arg Lys Val Thr Ala Gly Asp Met Gly Ser 275 280 285 Tyr Thr Cys Val Ala Glu Asn Met Val Gly Lys Ala Glu Ala Ser Ala 290 295 300 Thr Leu Thr Val Gln Val Gly Ser Glu Pro Pro His Phe Val Val Lys 305 310 315 320 Pro Arg Asp Gln Val Val Ala Leu Gly Arg Thr Val Thr Phe Gln Cys 325 330 335 Glu Ala Thr Gly Asn Pro Gln Pro Ala Ile Phe Trp Arg Arg Glu Gly 340 345 350 Ser Gln Asn Leu Leu Phe Ser Tyr Gln Pro Pro Gln Ser Ser Ser Arg 355 360 365 Phe Ser Val Ser Gln Thr Gly Asp Leu Thr Ile Thr Asn Val Gln Arg 370 375 380 Ser Asp Val Gly Tyr Tyr Ile Cys Gln Thr Leu Asn Val Ala Gly Ser 385 390 395 400 Ile Ile Thr Lys Ala Tyr Leu Glu Val Thr Asp Val Ile Ala Asp Arg 405 410 415 Pro Pro Pro Val Ile Arg Gln Gly Pro Val Asn Gln Thr Val Ala Val 420 425 430 Asp Gly Thr Phe Val Leu Ser Cys Val Ala Thr Gly Ser Pro Val Pro 435 440 445 Thr Ile Leu Trp Arg Lys Asp Gly Val Leu Val Ser Thr Gln Asp Ser 450 455 460 Arg Ile Lys Gln Leu Glu Asn Gly Val Leu Gln Ile Arg Tyr Ala Lys 465 470 475 480 Leu Gly Asp Thr Gly Arg Tyr Thr Cys Ile Ala Ser Thr Pro Ser Gly 485 490 495 Glu Ala Thr Trp Ser Ala Tyr Ile Glu Val Gln Glu Phe Gly Val Pro 500 505 510 Val Gln Pro Pro Arg Pro Thr Asp Pro Asn Leu Ile Pro Ser Ala Pro 515 520 525 Ser Lys Pro Glu Val Thr Asp Val Ser Arg Asn Thr Val Thr Leu Ser 530 535 540 Trp Gln Pro Asn Leu Asn Ser Gly Ala Thr Pro Thr Ser Tyr Ile Ile 545 550 555 560 Glu Ala Phe Ser His Ala Ser Gly Ser Ser Trp Gln Thr Val Ala Glu 565 570 575 Asn Val Lys Thr Glu Thr Ser Ala Ile Lys Gly Leu Lys Pro Asn Ala 580 585 590 Ile Tyr Leu Phe Leu Val Arg Ala Ala Asn Ala Tyr Gly Ile Ser Asp 595 600 605 Pro Ser Gln Ile Ser Asp Pro Val Lys Thr Gln Asp Val Leu Pro Thr 610 615 620 Ser Gln Gly Val Asp His Lys Gln Val Gln Arg Glu Leu Gly Asn Ala 625 630 635 640 Val Leu His Leu His Asn Pro Thr Val Leu Ser Ser Ser Ser Ile Glu 645 650 655 Val His Trp Thr Val Asp Gln Gln Ser Gln Tyr Ile Gln Gly Tyr Lys 660 665 670 Ile Leu Tyr Arg Pro Ser Gly Ala Asn His Gly Glu Ser Asp Trp Leu 675 680 685 Val Phe Glu Val Arg Thr Pro Ala Lys Asn Ser Val Val Ile Pro Asp 690 695 700 Leu Arg Lys Gly Val Asn Tyr Glu Ile Lys Ala Arg Pro Phe Phe Asn 705 710 715 720 Glu Phe Gln Gly Ala Asp Ser Glu Ile Lys Phe Ala Lys Thr Leu Glu 725 730 735 Glu Ala Pro Ser Ala Pro Pro Gln Gly Val Thr Val Ser Lys Asn Asp 740 745 750 Gly Asn Gly Thr Ala Ile Leu Val Ser Trp Gln Pro Pro Pro Glu Asp 755 760 765 Thr Gln Asn Gly Met Val Gln Glu Tyr Lys Val Trp Cys Leu Gly Asn 770 775 780 Glu Thr Arg Tyr His Ile Asn Lys Thr Val Asp Gly Ser Thr Phe Ser 785 790 795 800 Val Val Ile Pro Phe Leu Val Pro Gly Ile Arg Tyr Ser Val Glu Val 805 810 815 Ala Ala Ser Thr Gly Ala Gly Ser Gly Val Lys Ser Glu Pro Gln Phe 820 825 830 Ile Gln Leu Asp Ala His Gly Asn Pro Val Ser Pro Glu Asp Gln Val 835 840 845 Ser Leu Ala Gln Gln Ile Ser Asp Val Val Lys Gln Pro Ala Phe Ile 850 855 860 Ala Gly Ile Gly Ala Ala Cys Trp Ile Ile Leu Met Val Phe Ser Ile 865 870 875 880 Trp Leu Tyr Arg His Arg Lys Lys Arg Asn Gly Leu Thr Ser Thr Tyr 885 890 895 Ala Gly Ile Arg Lys Val Thr Tyr Gln Arg Gly Gly Glu Ala Val Ser 900 905 910 Ser Gly Gly Arg Pro Gly Leu Leu Asn Ile Ser Glu Pro Ala Ala Gln 915 920 925 Pro Trp Leu Ala Asp Thr Trp Pro Asn Thr Gly Asn Asn His Asn Asp 930 935 940 Cys Ser Ile Ser Cys Cys Thr Ala Gly Asn Gly Asn Ser Asp Ser Asn 945 950 955 960 Leu Thr Thr Tyr Ser Arg Pro Gly Gln Pro Thr Pro Tyr Ala Thr Thr 965 970 975 Gln Leu Ile Gln Ser Asn Leu Ser Asn Asn Met Asn Asn Gly Ser Gly 980 985 990 Asp Ser Gly Glu Lys His Trp Lys Pro Leu Gly Gln Gln Lys Gln Glu 995 1000 1005 Val Ala Pro Val Gln Tyr Asn Ile Val Glu Gln Asn Lys Leu Asn Lys 1010 1015 1020 Asp Tyr Arg Ala Asn Asp Thr Val Pro Pro Thr Ile Pro Tyr Asn Gln 1025 1030 1035 1040 Ser Tyr Asp Gln Asn Thr Gly Gly Ser Tyr Asn Ser Ser Asp Arg Gly 1045 1050 1055 Ser Ser Thr Ser Gly Ser Gln Gly His Lys Lys Gly Ala Arg Thr Pro 1060 1065 1070 Lys Val Pro Lys Gln Gly Gly Met Asn Trp Ala Asp Leu Leu Pro Pro 1075 1080 1085 Pro Pro Ala His Pro Pro Pro His Ser Asn Ser Glu Glu Tyr Asn Ile 1090 1095 1100 Ser Val Asp Glu Ser Tyr Asp Gln Glu Met Pro Cys Pro Val Pro Pro 1105 1110 1115 1120 Ala Arg Met Tyr Leu Gln Gln Asp Glu Leu Glu Glu Glu Glu Asp Glu 1125 1130 1135 Arg Gly Pro Thr Pro Pro Val Arg Gly Ala Ala Ser Ser Pro Ala Ala 1140 1145 1150 Val Ser Tyr Ser His Gln Ser Thr Ala Thr Leu Thr Pro Ser Pro Gln 1155 1160 1165 Glu Glu Leu Gln Pro Met Leu Gln Asp Cys Pro Glu Glu Thr Gly His 1170 1175 1180 Met Gln His Gln Pro Asp Arg Arg Arg Gln Pro Val Ser Pro Pro Pro 1185 1190 1195 1200 Pro Pro Arg Pro Ile Ser Pro Pro His Thr Tyr Gly Tyr Ile Ser Gly 1205 1210 1215 Pro Leu Val Ser Asp Met Asp Thr Asp Ala Pro Glu Glu Glu Glu Asp 1220 1225 1230 Glu Ala Asp Met Glu Val Ala Lys Met Gln Thr Arg Arg Leu Leu Leu 1235 1240 1245 Arg Gly Leu Glu Gln Thr Pro Ala Ser Ser Val Gly Asp Leu Glu Ser 1250 1255 1260 Ser Val Thr Gly Ser Met Ile Asn Gly Trp Gly Ser Ala Ser Glu Glu 1265 1270 1275 1280 Asp Asn Ile Ser Ser Gly Arg Ser Ser Val Ser Ser Ser Asp Gly Ser 1285 1290 1295 Phe Phe Thr Asp Ala Asp Phe Ala Gln Ala Val Ala Ala Ala Ala Glu 1300 1305 1310 Tyr Ala Gly Leu Lys Val Ala Arg Arg Gln Met Gln Asp Ala Ala Gly 1315 1320 1325 Arg Arg His Phe His Ala Ser Gln Cys Pro Arg Pro Thr Ser Pro Val 1330 1335 1340 Ser Thr Asp Ser Asn Met Ser Ala Ala Val Met Gln Lys Thr Arg Pro 1345 1350 1355 1360 Ala Lys Lys Leu Lys His Gln Pro Gly His Leu Arg Arg Glu Thr Tyr 1365 1370 1375 Thr Asp Asp Leu Pro Pro Pro Pro Val Pro Pro Pro Ala Ile Lys Ser 1380 1385 1390 Pro Thr Ala Gln Ser Lys Thr Gln Leu Glu Val Arg Pro Val Val Val 1395 1400 1405 Pro Lys Leu Pro Ser Met Asp Ala Arg Thr Asp Arg Ser Ser Asp Arg 1410 1415 1420 Lys Gly Ser Ser Tyr Lys Gly Arg Glu Val Leu Asp Gly Arg Gln Val 1425 1430 1435 1440 Val Asp Met Arg Thr Asn Pro Gly Asp Pro Arg Glu Ala Gln Glu Gln 1445 1450 1455 Gln Asn Asp Gly Lys Gly Arg Gly Asn Lys Ala Ala Lys Arg Asp Leu 1460 1465 1470 Pro Pro Ala Lys Thr His Leu Ile Gln Glu Asp Ile Leu Pro Tyr Cys 1475 1480 1485 Arg Pro Thr Phe Pro Thr Ser Asn Asn Pro Arg Asp Pro Ser Ser Ser 1490 1495 1500 Ser Ser Met Ser Ser Arg Gly Ser Gly Ser Arg Gln Arg Glu Gln Ala 1505 1510 1515 1520 Asn Val Gly Arg Arg Asn Ile Ala Glu Met Gln Val Leu Gly Gly Tyr 1525 1530 1535 Glu Arg Gly Glu Asp Asn Asn Glu Glu Leu Glu Glu Thr Glu Ser 1540 1545 1550 <210> 10 <211> 6895 <212> DNA <213> Homo sapiens roundabout, axon guidance receptor, homolog 1 (Drosophila) (ROBO1) <400> 10 cccgacttca ctctctccct atttccccac tcttaggttt aaaagtctgt cacctttcgc 60 ttggtttaaa ctcggaaagg tctcagtgca cagcaaagtt gcagggctgc gtctgcacta 120 cggagcctct agattgctga aaacagtctt atggaaggat aacacattgt ctgtcactgg 180 ctggttgtaa tgcaaggaag ggacaaagat gaaatggaaa catgttcctt ttttggtcat 240 gatatcactc ctcagcttat ccccaaatca cctgtttctg gcccagctta ttccagaccc 300 tgaagatgta gagaggggga acgaccacgg gacgccaatc cccacctctg ataacgatga 360 caattcgctg ggctatacag gctcccgtct tcgtcaggaa gattttccac ctcgcattgt 420 tgaacaccct tcagacctga ttgtctcaaa aggagaacct gcaactttga actgcaaagc 480 tgaaggccgc cccacaccca ctattgaatg gtacaaaggg ggagagagag tggagacaga 540 caaagatgac cctcgctcac accgaatgtt gctgccgagt ggatctttat ttttcttacg 600 tatagtacat ggacggaaaa gtagacctga tgaaggagtc tatgtctgtg tagcaaggaa 660 ttaccttgga gaggctgtga gccacaatgc atcgctggaa gtagccatac ttcgggatga 720 cttcagacaa aacccttcgg atgtcatggt tgcagtagga gagcctgcag taatggaatg 780 ccaacctcca cgaggccatc ctgagcccac catttcatgg aagaaagatg gctctccact 840 ggatgataaa gatgaaagaa taactatacg aggaggaaag ctcatgatca cttacacccg 900 taaaagtgac gctggcaaat atgtttgtgt tggtaccaat atggttgggg aacgtgagag 960 tgaagtagcc gagctgactg tcttagagag accatcattt gtgaagagac ccagtaactt 1020 ggcagtaact gtggatgaca gtgcagaatt taaatgtgag gcccgaggtg accctgtacc 1080 tacagtacga tggaggaaag atgatggaga gctgcccaaa tccagatatg aaatccgaga 1140 tgatcatacc ttgaaaatta ggaaggtgac agctggtgac atgggttcat acacttgtgt 1200 tgcagaaaat atggtgggca aagctgaagc atctgctact ctgactgttc aagaacctcc 1260 acattttgtt gtgaaacccc gtgaccaggt tgttgctttg ggacggactg taacttttca 1320 gtgtgaagca accggaaatc ctcaaccagc tattttctgg aggagagaag ggagtcagaa 1380 tctacttttc tcatatcaac caccacagtc atccagccga ttttcagtct cccagactgg 1440 cgacctcaca attactaatg tccagcgatc tgatgttggt tattacatct gccagacttt 1500 aaatgttgct ggaagcatca tcacaaaggc atatttggaa gttacagatg tgattgcaga 1560 tcggcctccc ccagttattc gacaaggtcc tgtgaatcag actgtagccg tggatggcac 1620 tttcgtcctc agctgtgtgg ccacaggcag tccagtgccc accattctgt ggagaaagga 1680 tggagtcctc gtttcaaccc aagactctcg aatcaaacag ttggagaatg gagtactgca 1740 gatccgatat gctaagctgg gtgatactgg tcggtacacc tgcattgcat caacccccag 1800 tggtgaagca acatggagtg cttacattga agttcaagaa tttggagttc cagttcagcc 1860 tccaagacct actgacccaa atttaatccc tagtgcccca tcaaaacctg aagtgacaga 1920 tgtcagcaga aatacagtca cattatcgtg gcaaccaaat ttgaattcag gagcaactcc 1980 aacatcttat attatagaag ccttcagcca tgcatctggt agcagctggc agaccgtagc 2040 agagaatgtg aaaacagaaa catctgccat taaaggactc aaacctaatg caatttacct 2100 tttccttgtg agggcagcta atgcatatgg aattagtgat ccaagccaaa tatcagatcc 2160 agtgaaaaca caagatgtcc taccaacaag tcagggggtg gaccacaagc aggtccagag 2220 agagctggga aatgctgttc tgcacctcca caaccccacc gtcctttctt cctcttccat 2280 cgaagtgcac tggacagtag atcaacagtc tcagtatata caaggatata aaattctcta 2340 tcggccatct ggagccaacc acggagaatc agactggtta gtttttgaag tgaggacgcc 2400 agccaaaaac agtgtggtaa tccctgatct cagaaaggga gtcaactatg aaattaaggc 2460 tcgccctttt tttaatgaat ttcaaggagc agatagtgaa atcaagtttg ccaaaaccct 2520 ggaagaagca cccagtgccc caccccaagg tgtaactgta tccaagaatg atggaaacgg 2580 aactgcaatt ctagttagtt ggcagccacc tccagaagac actcaaaatg gaatggtcca 2640 agagtataag gtttggtgtc tgggcaatga aactcgatac cacatcaaca aaacagtgga 2700 tggttccacc ttttccgtgg tcattccctt tcttgttcct ggaatccgat acagtgtgga 2760 agtggcagcc agcactgggg ctgggtctgg ggtaaagagt gagcctcagt tcatccagct 2820 ggatgcccat ggaaaccctg tgtcacctga ggaccaagtc agcctcgctc agcagatttc 2880 agatgtggtg aagcagccgg ccttcatagc aggtattgga gcagcctgtt ggatcatcct 2940 catggtcttc agcatctggc tttatcgaca ccgcaagaag agaaacggac ttactagtac 3000 ctacgcgggt atcagaaaag tcccgtcttt taccttcaca ccaacagtaa cttaccagag 3060 aggaggcgaa gctgtcagca gtggagggag gcctggactt ctcaacatca gtgaacctgc 3120 cgcgcagcca tggctggcag acacgtggcc taatactggc aacaaccaca atgactgctc 3180 catcagctgc tgcacggcag gcaatggaaa cagcgacagc aacctcacta cctacagtcg 3240 cccagctgat tgtatagcaa attataacaa ccaactggat aacaaacaaa caaatctgat 3300 gctccctgag tcaactgttt atggtgatgt ggaccttagt aacaaaatca atgagatgaa 3360 aaccttcaat agcccaaatc tgaaggatgg gcgttttgtc aatccatcag ggcagcctac 3420 tccttacgcc accactcagc tcatccagtc aaacctcagc aacaacatga acaatggcag 3480 cggggactct ggcgagaagc actggaaacc actgggacag cagaaacaag aagtggcacc 3540 agttcagtac aacatcgtgg agcaaaacaa gctgaacaaa gattatcgag caaatgacac 3600 agttcctcca actatcccat acaaccaatc atacgaccag aacacaggag gatcctacaa 3660 cagctcagac cggggcagta gtacatctgg gagtcagggg cacaagaaag gggcaagaac 3720 acccaaggta ccaaaacagg gtggcatgaa ctgggcagac ctgcttcctc ctcccccagc 3780 acatcctcct ccacacagca atagcgaaga gtacaacatt tctgtagatg aaagctatga 3840 ccaagaaatg ccatgtcccg tgccaccagc aaggatgtat ttgcaacaag atgaattaga 3900 agaggaggaa gatgaacgag gccccactcc ccctgttcgg ggagcagctt cttctccagc 3960 tgccgtgtcc tatagccatc agtccactgc cactctgact ccctccccac aggaagaact 4020 ccagcccatg ttacaggatt gtccagagga gactggccac atgcagcacc agcccgacag 4080 gagacggcag cctgtgagtc ctcctccacc accacggccg atctcccctc cacataccta 4140 tggctacatt tcaggacccc tggtctcaga tatggatacg gatgcgccag aagaggaaga 4200 agacgaagcc gacatggagg tagccaagat gcaaaccaga aggcttttgt tacgtgggct 4260 tgagcagaca cctgcctcca gtgttgggga cctggagagc tctgtcacgg ggtccatgat 4320 caacggctgg ggctcagcct cagaggagga caacatttcc agcggacgct ccagtgttag 4380 ttcttcggac ggctcctttt tcactgatgc tgactttgcc caggcagtcg cagcagcggc 4440 agagtatgct ggtctgaaag tagcacgacg gcaaatgcag gatgctgctg gccgtcgaca 4500 ttttcatgcg tctcagtgcc ctaggcccac aagtcccgtg tctacagaca gcaacatgag 4560 tgccgccgta atgcagaaaa ccagaccagc caagaaactg aaacaccagc caggacatct 4620 gcgcagagaa acctacacag atgatcttcc accacctcct gtgccgccac ctgctataaa 4680 gtcacctact gcccaatcca agacacagct ggaagtacga cctgtagtgg tgccaaaact 4740 cccttctatg gatgcaagaa cagacagatc atcagacaga aaaggaagca gttacaaggg 4800 gagagaagtg ttggatggaa gacaggttgt tgacatgcga acaaatccag gtgatcccag 4860 agaagcacag gaacagcaaa atgacgggaa aggacgtgga aacaaggcag caaaacgaga 4920 ccttccacca gcaaagactc atctcatcca agaggatatt ctaccttatt gtagacctac 4980 ttttccaaca tcaaataatc ccagagatcc cagttcctca agctcaatgt catcaagagg 5040 atcaggaagc agacaaagag aacaagcaaa tgtaggtcga agaaatattg cagaaatgca 5100 ggtacttgga ggatatgaaa gaggagaaga taataatgaa gaattagagg aaactgaaag 5160 ctgaagacaa ccaagaggct tatgagatct aatgtgaaaa tcatcactca agatgcctcc 5220 tgtcagatga cacatgacgc cagataaaat gttcagtgca atcagagtgt acaaattgtc 5280 gtttttattc ctcttattgg gatatcattt taaaaacttt attgggtttt tattgttgtt 5340 gtttgatccc taaccctaca aagagccttc ctattcccct cgctgttgga gcaaaccatt 5400 ataccttact tccagcaagc aaagtgcttt gacttcttgc ttcagtcatc agccagcaag 5460 agggaacaaa actgttcttt tgcattttgc cgctgagata tggcattgca ctgcttatat 5520 gccaagctaa tttatagcaa gatattgatc aaatatagaa agttgatatt caacctcaca 5580 agggctctca aagtataatc tttctatagc caactgctaa tgcaaattaa aacatatttc 5640 attttaacat gatttcaaaa tcagtttttc atactaccct ttgctggaag aaactaaaaa 5700 tatagcaaat gcagaaccac aaacaattcg aatggggtag aaacattgta aatatttact 5760 ctttgcaaac cctggtggta ttttattttg gcttcatttc aatcattgaa gtatattctt 5820 attggaaatg tacttttgga taagtagggc taagccagtt ggatctctgg ttgtctagtc 5880 attgtcataa gtaaacctag taaaaccttg ttctattttt caatcatcaa aaagtaatta 5940 taaatacgta ttacaaacaa gtggatgttt ttaatgacca attgagtaag aacatccctg 6000 tcttaactgg cctaaatttc ttctggtagt gtcagttcaa ctttcagaag tgccacttaa 6060 ggaagtttga tttttgtttt tgtaatgcac tgtttttaat ctctctctct tttttttttt 6120 ttttttggtt ttaaaagcac aatcactaaa ctttatttgt aaaccattgt aactattaac 6180 cttttttgtc ttattgaaaa aaaaaatgtt gagaagcgtt tttaacctgt tttgttaatg 6240 ctctatgttt gtatttggaa tatttgaata atgacagatg gtgaagtaac atgcatactt 6300 tattgtgggc catgaaccaa atggttctta cttttcctgg acttaaagaa aaaaagaggt 6360 ttaagtttgt tgtggccaat gtcgaaacct acaagatttc cttaaaatct ctaatagagg 6420 cattacttgc tttcaattga caaatgatgc cctctgacta gtagatttct atgatccttt 6480 tttgtcattt tatgaatatc attgatttta taattggtgc tatttgaaga aaaaaatgta 6540 catttattca tagatagata agtatcaggt ctgaccccag tggaaaacaa agccaaacaa 6600 aactgaacca caaaaaaaaa ggctggtgtt caccaaaacc aaacttgttc atttagataa 6660 tttgaaaaag ttccatagaa aaggcgtgca gtactaaggg aacaatccat gtgattaatg 6720 ttttcattat gttcatgtaa gaagcccctt atttttagcc ataattttgc atactgaaaa 6780 tccaataatc agaaaagtaa ttttgtcaca ttatttatta aaaatgttct caaatacata 6840 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaa 6895 <210> 11 <211> 7550 <212> DNA <213> Homo sapiens roundabout, axon guidance receptor, homolog 1 (Drosophila) (ROBO1) <400> 11 aattgagctg gagaggaggc agcgtgagag cagaaacttc agacgccgct gatccgggag 60 gagctggggt gagccgcggc ggccgtctct cccacccgca gcagcatcct ctctgccctt 120 ctctgccacc ccggggagag ccgggagctg cctctttaca gcttccacga gccaggggtg 180 caggcagctg cccccaggaa gtttgggctt ctgcgtagtt taggggtgcc tgcgagcgcc 240 ccagagggcg aggggccgag ggcgatgttg ggcgccgcgc ggggctgggg gcgcccagaa 300 gacgtgcgag tgtccgcggt cctgctgctg tctccagtac cctccgcatc ccccaagtga 360 tgggaacaag ggcccgccca ggcagccgct gtcgccgcac cgccccctcg ctcgctctct 420 gcgcgcggag tcacccagtc acactcccgg caccccgagc ccttcctccg gagctgctgc 480 ttctactttg gctgctatcg ccgccgccgc gggtggcccg ctgctgactg ggctcgccgg 540 gagacggaga agcacttttt ggccctccct cagcagctct cacaccccaa ctttgccgcc 600 gccgccgcgc ctgccctcgc agcggcgctc ggccgcacat tgtgggggcg cacgccggga 660 ggctccgcaa gaccgtggag gcaggaaacg gcactactgc gcttctgcct cggctctttg 720 ttgttcgctt tggatggttc ttgaaagtgt ctgagcctcc tcggaaatcc tggggccgga 780 gaagacaaac cttggaattc ttcctctgca aaagtctctg agatactgac aagcgtccgg 840 aaaggtcgac gagtaattgc cctgaaaact cttggctaat tgacccacgt tgcttatatt 900 aagcctttgt gtgtggtgtg tggcttcata catttgggga ccctatttcc actccctcct 960 cttggcatga gactgtatac aggatccacc cgaggacaat gattgcggag cccgctcact 1020 tttacctgtt tggattaata tgtctctgtt caggctcccg tcttcgtcag gaagattttc 1080 cacctcgcat tgttgaacac ccttcagacc tgattgtctc aaaaggagaa cctgcaactt 1140 tgaactgcaa agctgaaggc cgccccacac ccactattga atggtacaaa gggggagaga 1200 gagtggagac agacaaagat gaccctcgct cacaccgaat gttgctgccg agtggatctt 1260 tatttttctt acgtatagta catggacgga aaagtagacc tgatgaagga gtctatgtct 1320 gtgtagcaag gaattacctt ggagaggctg tgagccacaa tgcatcgctg gaagtagcca 1380 tacttcggga tgacttcaga caaaaccctt cggatgtcat ggttgcagta ggagagcctg 1440 cagtaatgga atgccaacct ccacgaggcc atcctgagcc caccatttca tggaagaaag 1500 atggctctcc actggatgat aaagatgaaa gaataactat acgaggagga aagctcatga 1560 tcacttacac ccgtaaaagt gacgctggca aatatgtttg tgttggtacc aatatggttg 1620 gggaacgtga gagtgaagta gccgagctga ctgtcttaga gagaccatca tttgtgaaga 1680 gacccagtaa cttggcagta actgtggatg acagtgcaga atttaaatgt gaggcccgag 1740 gtgaccctgt acctacagta cgatggagga aagatgatgg agagctgccc aaatccagat 1800 atgaaatccg agatgatcat accttgaaaa ttaggaaggt gacagctggt gacatgggtt 1860 catacacttg tgttgcagaa aatatggtgg gcaaagctga agcatctgct actctgactg 1920 ttcaagttgg gtctgaacct ccacattttg ttgtgaaacc ccgtgaccag gttgttgctt 1980 tgggacggac tgtaactttt cagtgtgaag caaccggaaa tcctcaacca gctattttct 2040 ggaggagaga agggagtcag aatctacttt tctcatatca accaccacag tcatccagcc 2100 gattttcagt ctcccagact ggcgacctca caattactaa tgtccagcga tctgatgttg 2160 gttattacat ctgccagact ttaaatgttg ctggaagcat catcacaaag gcatatttgg 2220 aagttacaga tgtgattgca gatcggcctc ccccagttat tcgacaaggt cctgtgaatc 2280 agactgtagc cgtggatggc actttcgtcc tcagctgtgt ggccacaggc agtccagtgc 2340 ccaccattct gtggagaaag gatggagtcc tcgtttcaac ccaagactct cgaatcaaac 2400 agttggagaa tggagtactg cagatccgat atgctaagct gggtgatact ggtcggtaca 2460 cctgcattgc atcaaccccc agtggtgaag caacatggag tgcttacatt gaagttcaag 2520 aatttggagt tccagttcag cctccaagac ctactgaccc aaatttaatc cctagtgccc 2580 catcaaaacc tgaagtgaca gatgtcagca gaaatacagt cacattatcg tggcaaccaa 2640 atttgaattc aggagcaact ccaacatctt atattataga agccttcagc catgcatctg 2700 gtagcagctg gcagaccgta gcagagaatg tgaaaacaga aacatctgcc attaaaggac 2760 tcaaacctaa tgcaatttac cttttccttg tgagggcagc taatgcatat ggaattagtg 2820 atccaagcca aatatcagat ccagtgaaaa cacaagatgt cctaccaaca agtcaggggg 2880 tggaccacaa gcaggtccag agagagctgg gaaatgctgt tctgcacctc cacaacccca 2940 ccgtcctttc ttcctcttcc atcgaagtgc actggacagt agatcaacag tctcagtata 3000 tacaaggata taaaattctc tatcggccat ctggagccaa ccacggagaa tcagactggt 3060 tagtttttga agtgaggacg ccagccaaaa acagtgtggt aatccctgat ctcagaaagg 3120 gagtcaacta tgaaattaag gctcgccctt tttttaatga atttcaagga gcagatagtg 3180 aaatcaagtt tgccaaaacc ctggaagaag cacccagtgc cccaccccaa ggtgtaactg 3240 tatccaagaa tgatggaaac ggaactgcaa ttctagttag ttggcagcca cctccagaag 3300 acactcaaaa tggaatggtc caagagtata aggtttggtg tctgggcaat gaaactcgat 3360 accacatcaa caaaacagtg gatggttcca ccttttccgt ggtcattccc tttcttgttc 3420 ctggaatccg atacagtgtg gaagtggcag ccagcactgg ggctgggtct ggggtaaaga 3480 gtgagcctca gttcatccag ctggatgccc atggaaaccc tgtgtcacct gaggaccaag 3540 tcagcctcgc tcagcagatt tcagatgtgg tgaagcagcc ggccttcata gcaggtattg 3600 gagcagcctg ttggatcatc ctcatggtct tcagcatctg gctttatcga caccgcaaga 3660 agagaaacgg acttactagt acctacgcgg gtatcagaaa agtaacttac cagagaggag 3720 gcgaagctgt cagcagtgga gggaggcctg gacttctcaa catcagtgaa cctgccgcgc 3780 agccatggct ggcagacacg tggcctaata ctggcaacaa ccacaatgac tgctccatca 3840 gctgctgcac ggcaggcaat ggaaacagcg acagcaacct cactacctac agtcgcccag 3900 ctgattgtat agcaaattat aacaaccaac tggataacaa acaaacaaat ctgatgctcc 3960 ctgagtcaac tgtttatggt gatgtggacc ttagtaacaa aatcaatgag atgaaaacct 4020 tcaatagccc aaatctgaag gatgggcgtt ttgtcaatcc atcagggcag cctactcctt 4080 acgccaccac tcagctcatc cagtcaaacc tcagcaacaa catgaacaat ggcagcgggg 4140 actctggcga gaagcactgg aaaccactgg gacagcagaa acaagaagtg gcaccagttc 4200 agtacaacat cgtggagcaa aacaagctga acaaagatta tcgagcaaat gacacagttc 4260 ctccaactat cccatacaac caatcatacg accagaacac aggaggatcc tacaacagct 4320 cagaccgggg cagtagtaca tctgggagtc aggggcacaa gaaaggggca agaacaccca 4380 aggtaccaaa acagggtggc atgaactggg cagacctgct tcctcctccc ccagcacatc 4440 ctcctccaca cagcaatagc gaagagtaca acatttctgt agatgaaagc tatgaccaag 4500 aaatgccatg tcccgtgcca ccagcaagga tgtatttgca acaagatgaa ttagaagagg 4560 aggaagatga acgaggcccc actccccctg ttcggggagc agcttcttct ccagctgccg 4620 tgtcctatag ccatcagtcc actgccactc tgactccctc cccacaggaa gaactccagc 4680 ccatgttaca ggattgtcca gaggagactg gccacatgca gcaccagccc gacaggagac 4740 ggcagcctgt gagtcctcct ccaccaccac ggccgatctc ccctccacat acctatggct 4800 acatttcagg acccctggtc tcagatatgg atacggatgc gccagaagag gaagaagacg 4860 aagccgacat ggaggtagcc aagatgcaaa ccagaaggct tttgttacgt gggcttgagc 4920 agacacctgc ctccagtgtt ggggacctgg agagctctgt cacggggtcc atgatcaacg 4980 gctggggctc agcctcagag gaggacaaca tttccagcgg acgctccagt gttagttctt 5040 cggacggctc ctttttcact gatgctgact ttgcccaggc agtcgcagca gcggcagagt 5100 atgctggtct gaaagtagca cgacggcaaa tgcaggatgc tgctggccgt cgacattttc 5160 atgcgtctca gtgccctagg cccacaagtc ccgtgtctac agacagcaac atgagtgccg 5220 ccgtaatgca gaaaaccaga ccagccaaga aactgaaaca ccagccagga catctgcgca 5280 gagaaaccta cacagatgat cttccaccac ctcctgtgcc gccacctgct ataaagtcac 5340 ctactgccca atccaagaca cagctggaag tacgacctgt agtggtgcca aaactccctt 5400 ctatggatgc aagaacagac agatcatcag acagaaaagg aagcagttac aaggggagag 5460 aagtgttgga tggaagacag gttgttgaca tgcgaacaaa tccaggtgat cccagagaag 5520 cacaggaaca gcaaaatgac gggaaaggac gtggaaacaa ggcagcaaaa cgagaccttc 5580 caccagcaaa gactcatctc atccaagagg atattctacc ttattgtaga cctacttttc 5640 caacatcaaa taatcccaga gatcccagtt cctcaagctc aatgtcatca agaggatcag 5700 gaagcagaca aagagaacaa gcaaatgtag gtcgaagaaa tattgcagaa atgcaggtac 5760 ttggaggata tgaaagagga gaagataata atgaagaatt agaggaaact gaaagctgaa 5820 gacaaccaag aggcttatga gatctaatgt gaaaatcatc actcaagatg cctcctgtca 5880 gatgacacat gacgccagat aaaatgttca gtgcaatcag agtgtacaaa ttgtcgtttt 5940 tattcctctt attgggatat cattttaaaa actttattgg gtttttattg ttgttgtttg 6000 atccctaacc ctacaaagag ccttcctatt cccctcgctg ttggagcaaa ccattatacc 6060 ttacttccag caagcaaagt gctttgactt cttgcttcag tcatcagcca gcaagaggga 6120 acaaaactgt tcttttgcat tttgccgctg agatatggca ttgcactgct tatatgccaa 6180 gctaatttat agcaagatat tgatcaaata tagaaagttg atattcaacc tcacaagggc 6240 tctcaaagta taatctttct atagccaact gctaatgcaa attaaaacat atttcatttt 6300 aacatgattt caaaatcagt ttttcatact accctttgct ggaagaaact aaaaatatag 6360 caaatgcaga accacaaaca attcgaatgg ggtagaaaca ttgtaaatat ttactctttg 6420 caaaccctgg tggtatttta ttttggcttc atttcaatca ttgaagtata ttcttattgg 6480 aaatgtactt ttggataagt agggctaagc cagttggatc tctggttgtc tagtcattgt 6540 cataagtaaa cctagtaaaa ccttgttcta tttttcaatc atcaaaaagt aattataaat 6600 acgtattaca aacaagtgga tgtttttaat gaccaattga gtaagaacat ccctgtctta 6660 actggcctaa atttcttctg gtagtgtcag ttcaactttc agaagtgcca cttaaggaag 6720 tttgattttt gtttttgtaa tgcactgttt ttaatctctc tctctttttt tttttttttt 6780 tggttttaaa agcacaatca ctaaacttta tttgtaaacc attgtaacta ttaacctttt 6840 ttgtcttatt gaaaaaaaaa atgttgagaa gcgtttttaa cctgttttgt taatgctcta 6900 tgtttgtatt tggaatattt gaataatgac agatggtgaa gtaacatgca tactttattg 6960 tgggccatga accaaatggt tcttactttt cctggactta aagaaaaaaa gaggtttaag 7020 tttgttgtgg ccaatgtcga aacctacaag atttccttaa aatctctaat agaggcatta 7080 cttgctttca attgacaaat gatgccctct gactagtaga tttctatgat ccttttttgt 7140 cattttatga atatcattga ttttataatt ggtgctattt gaagaaaaaa atgtacattt 7200 attcatagat agataagtat caggtctgac cccagtggaa aacaaagcca aacaaaactg 7260 aaccacaaaa aaaaaggctg gtgttcacca aaaccaaact tgttcattta gataatttga 7320 aaaagttcca tagaaaaggc gtgcagtact aagggaacaa tccatgtgat taatgttttc 7380 attatgttca tgtaagaagc cccttatttt tagccataat tttgcatact gaaaatccaa 7440 taatcagaaa agtaattttg tcacattatt tattaaaaat gttctcaaat acataaaaaa 7500 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7550 <210> 12 <211> 7385 <212> DNA <213> Homo sapiens roundabout, axon guidance receptor, homolog 1 (Drosophila) (ROBO1) <400> 12 aattgagctg gagaggaggc agcgtgagag cagaaacttc agacgccgct gatccgggag 60 gagctggggt gagccgcggc ggccgtctct cccacccgca gcagcatcct ctctgccctt 120 ctctgccacc ccggggagag ccgggagctg cctctttaca gcttccacga gccaggggtg 180 caggcagctg cccccaggaa gtttgggctt ctgcgtagtt taggggtgcc tgcgagcgcc 240 ccagagggcg aggggccgag ggcgatgttg ggcgccgcgc ggggctgggg gcgcccagaa 300 gacgtgcgag tgtccgcggt cctgctgctg tctccagtac cctccgcatc ccccaagtga 360 tgggaacaag ggcccgccca ggcagccgct gtcgccgcac cgccccctcg ctcgctctct 420 gcgcgcggag tcacccagtc acactcccgg caccccgagc ccttcctccg gagctgctgc 480 ttctactttg gctgctatcg ccgccgccgc gggtggcccg ctgctgactg ggctcgccgg 540 gagacggaga agcacttttt ggccctccct cagcagctct cacaccccaa ctttgccgcc 600 gccgccgcgc ctgccctcgc agcggcgctc ggccgcacat tgtgggggcg cacgccggga 660 ggctccgcaa gaccgtggag gcaggaaacg gcactactgc gcttctgcct cggctctttg 720 ttgttcgctt tggatggttc ttgaaagtgt ctgagcctcc tcggaaatcc tggggccgga 780 gaagacaaac cttggaattc ttcctctgca aaagtctctg agatactgac aagcgtccgg 840 aaaggtcgac gagtaattgc cctgaaaact cttggctaat tgacccacgt tgcttatatt 900 aagcctttgt gtgtggtgtg tggcttcata catttgggga ccctatttcc actccctcct 960 cttggcatga gactgtatac aggatccacc cgaggacaat gattgcggag cccgctcact 1020 tttacctgtt tggattaata tgtctctgtt caggctcccg tcttcgtcag gaagattttc 1080 cacctcgcat tgttgaacac ccttcagacc tgattgtctc aaaaggagaa cctgcaactt 1140 tgaactgcaa agctgaaggc cgccccacac ccactattga atggtacaaa gggggagaga 1200 gagtggagac agacaaagat gaccctcgct cacaccgaat gttgctgccg agtggatctt 1260 tatttttctt acgtatagta catggacgga aaagtagacc tgatgaagga gtctatgtct 1320 gtgtagcaag gaattacctt ggagaggctg tgagccacaa tgcatcgctg gaagtagcca 1380 tacttcggga tgacttcaga caaaaccctt cggatgtcat ggttgcagta ggagagcctg 1440 cagtaatgga atgccaacct ccacgaggcc atcctgagcc caccatttca tggaagaaag 1500 atggctctcc actggatgat aaagatgaaa gaataactat acgaggagga aagctcatga 1560 tcacttacac ccgtaaaagt gacgctggca aatatgtttg tgttggtacc aatatggttg 1620 gggaacgtga gagtgaagta gccgagctga ctgtcttaga gagaccatca tttgtgaaga 1680 gacccagtaa cttggcagta actgtggatg acagtgcaga atttaaatgt gaggcccgag 1740 gtgaccctgt acctacagta cgatggagga aagatgatgg agagctgccc aaatccagat 1800 atgaaatccg agatgatcat accttgaaaa ttaggaaggt gacagctggt gacatgggtt 1860 catacacttg tgttgcagaa aatatggtgg gcaaagctga agcatctgct actctgactg 1920 ttcaagttgg gtctgaacct ccacattttg ttgtgaaacc ccgtgaccag gttgttgctt 1980 tgggacggac tgtaactttt cagtgtgaag caaccggaaa tcctcaacca gctattttct 2040 ggaggagaga agggagtcag aatctacttt tctcatatca accaccacag tcatccagcc 2100 gattttcagt ctcccagact ggcgacctca caattactaa tgtccagcga tctgatgttg 2160 gttattacat ctgccagact ttaaatgttg ctggaagcat catcacaaag gcatatttgg 2220 aagttacaga tgtgattgca gatcggcctc ccccagttat tcgacaaggt cctgtgaatc 2280 agactgtagc cgtggatggc actttcgtcc tcagctgtgt ggccacaggc agtccagtgc 2340 ccaccattct gtggagaaag gatggagtcc tcgtttcaac ccaagactct cgaatcaaac 2400 agttggagaa tggagtactg cagatccgat atgctaagct gggtgatact ggtcggtaca 2460 cctgcattgc atcaaccccc agtggtgaag caacatggag tgcttacatt gaagttcaag 2520 aatttggagt tccagttcag cctccaagac ctactgaccc aaatttaatc cctagtgccc 2580 catcaaaacc tgaagtgaca gatgtcagca gaaatacagt cacattatcg tggcaaccaa 2640 atttgaattc aggagcaact ccaacatctt atattataga agccttcagc catgcatctg 2700 gtagcagctg gcagaccgta gcagagaatg tgaaaacaga aacatctgcc attaaaggac 2760 tcaaacctaa tgcaatttac cttttccttg tgagggcagc taatgcatat ggaattagtg 2820 atccaagcca aatatcagat ccagtgaaaa cacaagatgt cctaccaaca agtcaggggg 2880 tggaccacaa gcaggtccag agagagctgg gaaatgctgt tctgcacctc cacaacccca 2940 ccgtcctttc ttcctcttcc atcgaagtgc actggacagt agatcaacag tctcagtata 3000 tacaaggata taaaattctc tatcggccat ctggagccaa ccacggagaa tcagactggt 3060 tagtttttga agtgaggacg ccagccaaaa acagtgtggt aatccctgat ctcagaaagg 3120 gagtcaacta tgaaattaag gctcgccctt tttttaatga atttcaagga gcagatagtg 3180 aaatcaagtt tgccaaaacc ctggaagaag cacccagtgc cccaccccaa ggtgtaactg 3240 tatccaagaa tgatggaaac ggaactgcaa ttctagttag ttggcagcca cctccagaag 3300 acactcaaaa tggaatggtc caagagtata aggtttggtg tctgggcaat gaaactcgat 3360 accacatcaa caaaacagtg gatggttcca ccttttccgt ggtcattccc tttcttgttc 3420 ctggaatccg atacagtgtg gaagtggcag ccagcactgg ggctgggtct ggggtaaaga 3480 gtgagcctca gttcatccag ctggatgccc atggaaaccc tgtgtcacct gaggaccaag 3540 tcagcctcgc tcagcagatt tcagatgtgg tgaagcagcc ggccttcata gcaggtattg 3600 gagcagcctg ttggatcatc ctcatggtct tcagcatctg gctttatcga caccgcaaga 3660 agagaaacgg acttactagt acctacgcgg gtatcagaaa agtaacttac cagagaggag 3720 gcgaagctgt cagcagtgga gggaggcctg gacttctcaa catcagtgaa cctgccgcgc 3780 agccatggct ggcagacacg tggcctaata ctggcaacaa ccacaatgac tgctccatca 3840 gctgctgcac ggcaggcaat ggaaacagcg acagcaacct cactacctac agtcgcccag 3900 ggcagcctac tccttacgcc accactcagc tcatccagtc aaacctcagc aacaacatga 3960 acaatggcag cggggactct ggcgagaagc actggaaacc actgggacag cagaaacaag 4020 aagtggcacc agttcagtac aacatcgtgg agcaaaacaa gctgaacaaa gattatcgag 4080 caaatgacac agttcctcca actatcccat acaaccaatc atacgaccag aacacaggag 4140 gatcctacaa cagctcagac cggggcagta gtacatctgg gagtcagggg cacaagaaag 4200 gggcaagaac acccaaggta ccaaaacagg gtggcatgaa ctgggcagac ctgcttcctc 4260 ctcccccagc acatcctcct ccacacagca atagcgaaga gtacaacatt tctgtagatg 4320 aaagctatga ccaagaaatg ccatgtcccg tgccaccagc aaggatgtat ttgcaacaag 4380 atgaattaga agaggaggaa gatgaacgag gccccactcc ccctgttcgg ggagcagctt 4440 cttctccagc tgccgtgtcc tatagccatc agtccactgc cactctgact ccctccccac 4500 aggaagaact ccagcccatg ttacaggatt gtccagagga gactggccac atgcagcacc 4560 agcccgacag gagacggcag cctgtgagtc ctcctccacc accacggccg atctcccctc 4620 cacataccta tggctacatt tcaggacccc tggtctcaga tatggatacg gatgcgccag 4680 aagaggaaga agacgaagcc gacatggagg tagccaagat gcaaaccaga aggcttttgt 4740 tacgtgggct tgagcagaca cctgcctcca gtgttgggga cctggagagc tctgtcacgg 4800 ggtccatgat caacggctgg ggctcagcct cagaggagga caacatttcc agcggacgct 4860 ccagtgttag ttcttcggac ggctcctttt tcactgatgc tgactttgcc caggcagtcg 4920 cagcagcggc agagtatgct ggtctgaaag tagcacgacg gcaaatgcag gatgctgctg 4980 gccgtcgaca ttttcatgcg tctcagtgcc ctaggcccac aagtcccgtg tctacagaca 5040 gcaacatgag tgccgccgta atgcagaaaa ccagaccagc caagaaactg aaacaccagc 5100 caggacatct gcgcagagaa acctacacag atgatcttcc accacctcct gtgccgccac 5160 ctgctataaa gtcacctact gcccaatcca agacacagct ggaagtacga cctgtagtgg 5220 tgccaaaact cccttctatg gatgcaagaa cagacagatc atcagacaga aaaggaagca 5280 gttacaaggg gagagaagtg ttggatggaa gacaggttgt tgacatgcga acaaatccag 5340 gtgatcccag agaagcacag gaacagcaaa atgacgggaa aggacgtgga aacaaggcag 5400 caaaacgaga ccttccacca gcaaagactc atctcatcca agaggatatt ctaccttatt 5460 gtagacctac ttttccaaca tcaaataatc ccagagatcc cagttcctca agctcaatgt 5520 catcaagagg atcaggaagc agacaaagag aacaagcaaa tgtaggtcga agaaatattg 5580 cagaaatgca ggtacttgga ggatatgaaa gaggagaaga taataatgaa gaattagagg 5640 aaactgaaag ctgaagacaa ccaagaggct tatgagatct aatgtgaaaa tcatcactca 5700 agatgcctcc tgtcagatga cacatgacgc cagataaaat gttcagtgca atcagagtgt 5760 acaaattgtc gtttttattc ctcttattgg gatatcattt taaaaacttt attgggtttt 5820 tattgttgtt gtttgatccc taaccctaca aagagccttc ctattcccct cgctgttgga 5880 gcaaaccatt ataccttact tccagcaagc aaagtgcttt gacttcttgc ttcagtcatc 5940 agccagcaag agggaacaaa actgttcttt tgcattttgc cgctgagata tggcattgca 6000 ctgcttatat gccaagctaa tttatagcaa gatattgatc aaatatagaa agttgatatt 6060 caacctcaca agggctctca aagtataatc tttctatagc caactgctaa tgcaaattaa 6120 aacatatttc attttaacat gatttcaaaa tcagtttttc atactaccct ttgctggaag 6180 aaactaaaaa tatagcaaat gcagaaccac aaacaattcg aatggggtag aaacattgta 6240 aatatttact ctttgcaaac cctggtggta ttttattttg gcttcatttc aatcattgaa 6300 gtatattctt attggaaatg tacttttgga taagtagggc taagccagtt ggatctctgg 6360 ttgtctagtc attgtcataa gtaaacctag taaaaccttg ttctattttt caatcatcaa 6420 aaagtaatta taaatacgta ttacaaacaa gtggatgttt ttaatgacca attgagtaag 6480 aacatccctg tcttaactgg cctaaatttc ttctggtagt gtcagttcaa ctttcagaag 6540 tgccacttaa ggaagtttga tttttgtttt tgtaatgcac tgtttttaat ctctctctct 6600 tttttttttt ttttttggtt ttaaaagcac aatcactaaa ctttatttgt aaaccattgt 6660 aactattaac cttttttgtc ttattgaaaa aaaaaatgtt gagaagcgtt tttaacctgt 6720 tttgttaatg ctctatgttt gtatttggaa tatttgaata atgacagatg gtgaagtaac 6780 atgcatactt tattgtgggc catgaaccaa atggttctta cttttcctgg acttaaagaa 6840 aaaaagaggt ttaagtttgt tgtggccaat gtcgaaacct acaagatttc cttaaaatct 6900 ctaatagagg cattacttgc tttcaattga caaatgatgc cctctgacta gtagatttct 6960 atgatccttt tttgtcattt tatgaatatc attgatttta taattggtgc tatttgaaga 7020 aaaaaatgta catttattca tagatagata agtatcaggt ctgaccccag tggaaaacaa 7080 agccaaacaa aactgaacca caaaaaaaaa ggctggtgtt caccaaaacc aaacttgttc 7140 atttagataa tttgaaaaag ttccatagaa aaggcgtgca gtactaaggg aacaatccat 7200 gtgattaatg ttttcattat gttcatgtaa gaagcccctt atttttagcc ataattttgc 7260 atactgaaaa tccaataatc agaaaagtaa ttttgtcaca ttatttatta aaaatgttct 7320 caaatacata aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7380 aaaaa 7385 <110> THE ASAN FOUNDATION <120> A composition for preventing or inhibiting a metastasis of a primary thyroid cancer <130> PN140635 <160> 12 <170> Kopatentin 2.0 <210> 1 <211> 1529 <212> PRT <213> slit homolog 2 protein isoform 1 precursor [Homo sapiens] <400> 1 Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val 1 5 10 15 Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys 20 25 30 Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser 35 40 45 Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly 50 55 60 Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His 65 70 75 80 Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg 85 90 95 Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg 100 105 110 Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys 115 120 125 Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg 130 135 140 Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr 145 150 155 160 Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp 165 170 175 Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val 180 185 190 Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser 195 200 205 Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu 210 215 220 Arg Gln Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser 225 230 235 240 His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val 245 250 255 Cys Ser Gly His Gln Ser Phe Met Ala Pro Ser Cys Ser Val Leu His 260 265 270 Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val Asp Cys Arg Gly 275 280 285 Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu Thr Ile Thr Glu 290 295 300 Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro Pro Gly Ala Phe 305 310 315 320 Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser Asn Asn Gln Ile 325 330 335 Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg Ser Leu Asn Ser 340 345 350 Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro Lys Ser Leu Phe 355 360 365 Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn Ala Asn Lys Ile 370 375 380 Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His Asn Leu Asn Leu 385 390 395 400 Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala Lys Gly Thr Phe 405 410 415 Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala Gln Asn Pro Phe 420 425 430 Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr Leu His Thr Asn 435 440 445 Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro Arg Arg Leu Ala 450 455 460 Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe Arg Cys Ser Ala 465 470 475 480 Lys Glu Gln Tyr Phe Ile Pro Gly Thr Glu Asp Tyr Arg Ser Lys Leu 485 490 495 Ser Gly Asp Cys Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys 500 505 510 Glu Gly Thr Thr Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile Pro 515 520 525 Glu His Ile Pro Gln Tyr Thr Ala Glu Leu Arg Leu Asn Asn Asn Glu 530 535 540 Phe Thr Val Leu Glu Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu 545 550 555 560 Arg Lys Ile Asn Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly 565 570 575 Ala Phe Glu Gly Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn 580 585 590 Arg Leu Glu Asn Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu 595 600 605 Lys Thr Leu Met Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp 610 615 620 Ser Phe Ile Gly Leu Ser Ser Val Arg Leu Leu Ser Leu Tyr Asp Asn 625 630 635 640 Gln Ile Thr Thr Val Ala Pro Gly Ala Phe Asp Thr Leu His Ser Leu 645 650 655 Ser Thr Leu Asn Leu Leu Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu 660 665 670 Ala Trp Leu Gly Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn 675 680 685 Pro Arg Cys Gln Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp 690 695 700 Val Ala Ile Gln Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser 705 710 715 720 Cys Ser Pro Leu Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu Asp Thr 725 730 735 Val Val Arg Cys Ser Asn Lys Gly Leu Lys Val Leu Pro Lys Gly Ile 740 745 750 Pro Arg Asp Val Thr Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu 755 760 765 Val Pro Lys Glu Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu 770 775 780 Ser Asn Asn Arg Ile Ser Thr Leu Ser Asn Gln Ser Ser Phe Ser Asn Met 785 790 795 800 Thr Gln Leu Leu Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile 805 810 815 Pro Pro Arg Thr Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu 820 825 830 His Gly Asn Asp Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu 835 840 845 Ser Ala Leu Ser His Leu Ala Ile Gly Ala Asn Pro Leu Tyr Cys Asp 850 855 860 Cys Asn Met Gln Trp Leu Ser Asp Trp Val Lys Ser Glu Tyr Lys Glu 865 870 875 880 Pro Gly Ile Ala Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu 885 890 895 Leu Leu Thr Thr Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp 900 905 910 Val Asn Ile Leu Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys 915 920 925 Asn Asp Gly Thr Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr 930 935 940 Cys Pro Tyr Gly Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala 945 950 955 960 Cys Ile Ser Asn Pro Cys Lys His Gly Gly Thr Cys His Leu Lys Glu 965 970 975 Gly Glu Glu Asp Gly Phe Trp Cys Ile Cys Ala Asp Gly Phe Glu Gly 980 985 990 Glu Asn Cys Glu Val Asn Val Asp Asp Cys Glu Asp Asn Asp Cys Glu 995 1000 1005 Asn Asn Ser Thr Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu Cys 1010 1015 1020 Pro Pro Glu Tyr Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp Phe Cys 1025 1030 1035 1040 Ala Gln Asp Leu Asn Pro Cys Gln His Asp Ser Lys Cys Ile Leu Thr 1045 1050 1055 Pro Lys Gly Phe Lys Cys Asp Cys Thr Pro Gly Tyr Val Gly Glu His 1060 1065 1070 Cys Asp Ile Asp Phe Asp Asp Cys Gln Asp Asn Lys Cys Lys Asn Gly 1075 1080 1085 Ala His Cys Thr Asp Ala Val Asn Gly Tyr Thr Cys Ile Cys Pro Glu 1090 1095 1100 Gly Tyr Ser Gly Leu Phe Cys Glu Phe Ser Pro Pro Met Val Leu Pro 1105 1110 1115 1120 Arg Thr Ser Pro Cys Asp Asn Phe Asp Cys Gln Asn Gly Ala Gln Cys 1125 1130 1135 Ile Val Arg Ile Asn Glu Pro Ile Cys Gln Cys Leu Pro Gly Tyr Gln 1140 1145 1150 Gly Glu Lys Cys Glu Lys Leu Val Ser Val Asn Phe Ile Asn Lys Glu 1155 1160 1165 Ser Tyr Leu Gln Ile Ser Ser Ala Lys Val Arg Pro Gln Thr Asn Ile 1170 1175 1180 Thr Leu Gln Ile Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys 1185 1190 1195 1200 Gly Asp Lys Asp His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Arg 1205 1210 1215 Ala Ser Tyr Asp Thr Gly Ser His Pro Ala Ser Ala Ile Tyr Ser Val 1220 1225 1230 Glu Thr Ile Asn Asp Gly Asn Phe His Ile Val Glu Leu Leu Ala Leu 1235 1240 1245 Asp Gln Ser Leu Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile Ile 1250 1255 1260 Thr Asn Leu Ser Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro Leu Tyr 1265 1270 1275 1280 Val Gly Gly Met Pro Gly Lys Ser Asn Val Ala Ser Leu Arg Gln Ala 1285 1290 1295 Pro Gly Gln Asn Gly Thr Ser Phe His Gly Cys Ile Arg Asn Leu Tyr 1300 1305 1310 Ile Asn Ser Glu Leu Gln Asp Phe Gln Lys Val Pro Met Gln Thr Gly 1315 1320 1325 Ile Leu Pro Gly Cys Glu Pro Cys His Lys Lys Val Cys Ala His Gly 1330 1335 1340 Thr Cys Gln Pro Ser Ser Gln Ala Gly Phe Thr Cys Glu Cys Gln Glu 1345 1350 1355 1360 Gly Trp Met Gly Pro Leu Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu 1365 1370 1375 Gly Asn Lys Cys Val His Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser 1380 1385 1390 Tyr Ser Cys Lys Cys Leu Glu Gly His Gly Gly Val Leu Cys Asp Glu 1395 1400 1405 Glu Glu Asp Leu Phe Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly 1410 1415 1420 Lys Cys Arg Leu Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser 1425 1430 1435 1440 Gly Tyr Thr Gly Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu 1445 1450 1455 Arg Ile Arg Asp Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln 1460 1465 1470 Thr Thys Lys Lys Val Ser Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly 1475 1480 1485 Gly Gln Cys Cys Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe 1490 1495 1500 Glu Cys Thr Asp Gly Ser Ser Phe Val Asp Glu Val Glu Lys Val Val 1505 1510 1515 1520 Lys Cys Gly Cys Thr Arg Cys Val Ser 1525 <210> 2 <211> 1525 <212> PRT <213> slit homolog 2 protein isoform 2 precursor [Homo sapiens] <400> 2 Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val 1 5 10 15 Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys 20 25 30 Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser 35 40 45 Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly 50 55 60 Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His 65 70 75 80 Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg 85 90 95 Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg 100 105 110 Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys 115 120 125 Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg 130 135 140 Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr 145 150 155 160 Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp 165 170 175 Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val 180 185 190 Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser 195 200 205 Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu 210 215 220 Arg Gln Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser 225 230 235 240 His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val 245 250 255 Cys Ser Asp Glu Glu Glu Gly His Gln Ser Phe Met Ala Pro Ser Cys 260 265 270 Ser Val Leu His Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val 275 280 285 Asp Cys Arg Gly Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu 290 295 300 Thr Ile Thr Glu Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro 305 310 315 320 Pro Gly Ala Phe Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser 325 330 335 Asn Asn Gln Ile Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg 340 345 350 Ser Leu Asn Ser Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro 355 360 365 Lys Ser Leu Phe Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn 370 375 380 Ala Asn Lys Ile Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His 385 390 395 400 Asn Leu Asn Leu Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala 405 410 415 Lys Gly Thr Phe Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala 420 425 430 Gln Asn Pro Phe Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr 435 440 445 Leu His Thr Asn Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro 450 455 460 Arg Arg Leu Ala Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe 465 470 475 480 Arg Cys Ser Gly Thr Glu Asp Tyr Arg Ser Lys Leu Ser Gly Asp Cys 485 490 495 Phe Ala Asp Leu Ala Cys Pro Glu Lys Cys Arg Cys Glu Gly Thr Thr 500 505 510 Val Asp Cys Ser Asn Gln Lys Leu Asn Lys Ile Pro Glu His Ile Pro 515 520 525 Gln Tyr Thr Ala Glu Leu Arg Leu Asn Asn Asn Glu Phe Thr Val Leu 530 535 540 Glu Ala Thr Gly Ile Phe Lys Lys Leu Pro Gln Leu Arg Lys Ile Asn 545 550 555 560 Phe Ser Asn Asn Lys Ile Thr Asp Ile Glu Glu Gly Ala Phe Glu Gly 565 570 575 Ala Ser Gly Val Asn Glu Ile Leu Leu Thr Ser Asn Arg Leu Glu Asn 580 585 590 Val Gln His Lys Met Phe Lys Gly Leu Glu Ser Leu Lys Thr Leu Met 595 600 605 Leu Arg Ser Asn Arg Ile Thr Cys Val Gly Asn Asp Ser Phe Ile Gly 610 615 620 Leu Ser Ser Val Leu Leu Ser Leu Tyr Asp Asn Gln Ile Thr Thr 625 630 635 640 Val Ala Pro Gly Ala Phe Asp Thr Leu His Ser Leu Ser Thr Leu Asn 645 650 655 Leu Leu Ala Asn Pro Phe Asn Cys Asn Cys Tyr Leu Ala Trp Leu Gly 660 665 670 Glu Trp Leu Arg Lys Lys Arg Ile Val Thr Gly Asn Pro Arg Cys Gln 675 680 685 Lys Pro Tyr Phe Leu Lys Glu Ile Pro Ile Gln Asp Val Ala Ile Gln 690 695 700 Asp Phe Thr Cys Asp Asp Gly Asn Asp Asp Asn Ser Cys Ser Pro Leu 705 710 715 720 Ser Arg Cys Pro Thr Glu Cys Thr Cys Leu Asp Thr Val Val Arg Cys 725 730 735 Ser Asn Lys Gly Leu Lys Val Leu Pro Lys Gly Ile Pro Arg Asp Val 740 745 750 Thr Glu Leu Tyr Leu Asp Gly Asn Gln Phe Thr Leu Val Pro Lys Glu 755 760 765 Leu Ser Asn Tyr Lys His Leu Thr Leu Ile Asp Leu Ser Asn Asn Arg 770 775 780 Ile Ser Thr Leu Ser Asn Gln Ser Phe Ser Asn Met Thr Gln Leu Leu 785 790 795 800 Thr Leu Ile Leu Ser Tyr Asn Arg Leu Arg Cys Ile Pro Pro Arg Thr 805 810 815 Phe Asp Gly Leu Lys Ser Leu Arg Leu Leu Ser Leu His Gly Asn Asp 820 825 830 Ile Ser Val Val Pro Glu Gly Ala Phe Asn Asp Leu Ser Ala Leu Ser 835 840 845 His Leu Ala Ile Gly Ala Asn Pro Leu Tyr Cys Asp Cys Asn Met Gln 850 855 860 Trp Leu Ser Asp Trp Val Lys Ser Glu Tyr Lys Glu Pro Gly Ile Ala 865 870 875 880 Arg Cys Ala Gly Pro Gly Glu Met Ala Asp Lys Leu Leu Leu Thr Thr 885 890 895 Pro Ser Lys Lys Phe Thr Cys Gln Gly Pro Val Asp Val Asn Ile Leu 900 905 910 Ala Lys Cys Asn Pro Cys Leu Ser Asn Pro Cys Lys Asn Asp Gly Thr 915 920 925 Cys Asn Ser Asp Pro Val Asp Phe Tyr Arg Cys Thr Cys Pro Tyr Gly 930 935 940 Phe Lys Gly Gln Asp Cys Asp Val Pro Ile His Ala Cys Ile Ser Asn 945 950 955 960 Pro Cys Lys His Gly Gly Thr Cys His Leu Lys Glu Gly Glu Glu Asp 965 970 975 Gly Phe Trp Cys Ile Cys Ala Asp Gly Phe Glu Gly Glu Asn Cys Glu 980 985 990 Val Asn Val Asp Asp Cys Glu Asp Asn Asp Cys Glu Asn Asn Ser Thr 995 1000 1005 Cys Val Asp Gly Ile Asn Asn Tyr Thr Cys Leu Cys Pro Pro Glu Tyr 1010 1015 1020 Thr Gly Glu Leu Cys Glu Glu Lys Leu Asp Phe Cys Ala Gln Asp Leu 1025 1030 1035 1040 Asn Pro Cys Gln His Asp Ser Lys Cys Ile Leu Thr Pro Lys Gly Phe 1045 1050 1055 Lys Cys Asp Cys Thr Pro Gly Tyr Val Gly Glu His Cys Asp Ile Asp 1060 1065 1070 Phe Asp Asp Cys Gln Asp Asn Lys Cys Lys Asn Gly Ala His Cys Thr 1075 1080 1085 Asp Ala Val Asn Gly Tyr Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly 1090 1095 1100 Leu Phe Cys Glu Phe Ser Pro Pro Met Val Leu Pro Arg Thr Ser Pro 1105 1110 1115 1120 Cys Asp Asn Phe Asp Cys Gln Asn Gly Ala Gln Cys Ile Val Arg Ile 1125 1130 1135 Asn Glu Pro Ile Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys 1140 1145 1150 Glu Lys Leu Val Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln 1155 1160 1165 Ile Pro Ser Ala Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile 1170 1175 1180 Ala Thr Asp Glu Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys Asp 1185 1190 1195 1200 His Ile Ala Val Glu Leu Tyr Arg Gly Arg Val Val Ala Ser Tyr Asp 1205 1210 1215 Thr Gly Ser His Ala Ser Ala Ile Tyr Ser Val Glu Thr Ile Asn 1220 1225 1230 Asp Gly Asn Phe His Ile Val Glu Leu Leu Ala Leu Asp Gln Ser Leu 1235 1240 1245 Ser Leu Ser Val Asp Gly Gly Asn Pro Lys Ile Ile Thr Asn Leu Ser 1250 1255 1260 Lys Gln Ser Thr Leu Asn Phe Asp Ser Pro Leu Tyr Val Gly Gly Met 1265 1270 1275 1280 Pro Gly Lys Ser Asn Val Ala Ser Leu Arg Gln Ala Pro Gly Gln Asn 1285 1290 1295 Gly Thr Ser Phe His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu 1300 1305 1310 Leu Gln Asp Phe Gln Lys Val Pro Met Gln Thr Gly Ile Leu Pro Gly 1315 1320 1325 Cys Glu Pro Cys His Lys Lys Val Cys Ala His Gly Thr Cys Gln Pro 1330 1335 1340 Ser Ser Gln Ala Gly Phe Thr Cys Glu Cys Gln Glu Gly Trp Met Gly 1345 1350 1355 1360 Pro Leu Cys Asp Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys 1365 1370 1375 Val His Gly Thr Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys 1380 1385 1390 Cys Leu Glu Gly His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu 1395 1400 1405 Phe Asn Pro Cys Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu 1410 1415 1420 Ser Gly Leu Gly Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr Gly 1425 1430 1435 1440 Asp Ser Cys Asp Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp 1445 1450 1455 Tyr Tyr Gln Lys Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys 1460 1465 1470 Val Ser Arg Leu Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys 1475 1480 1485 Gly Pro Leu Arg Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp 1490 1495 1500 Gly Ser Ser Phe Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys 1505 1510 1515 1520 Thr Arg Cys Val Ser 1525 <210> 3 <211> 1521 <212> PRT <213> slit homolog 2 protein isoform 3 precursor [Homo sapiens] <400> 3 Met Arg Gly Val Gly Trp Gln Met Leu Ser Leu Ser Leu Gly Leu Val 1 5 10 15 Leu Ala Ile Leu Asn Lys Val Ala Pro Gln Ala Cys Pro Ala Gln Cys 20 25 30 Ser Cys Ser Gly Ser Thr Val Asp Cys His Gly Leu Ala Leu Arg Ser 35 40 45 Val Pro Arg Asn Ile Pro Arg Asn Thr Glu Arg Leu Asp Leu Asn Gly 50 55 60 Asn Asn Ile Thr Arg Ile Thr Lys Thr Asp Phe Ala Gly Leu Arg His 65 70 75 80 Leu Arg Val Leu Gln Leu Met Glu Asn Lys Ile Ser Thr Ile Glu Arg 85 90 95 Gly Ala Phe Gln Asp Leu Lys Glu Leu Glu Arg Leu Arg Leu Asn Arg 100 105 110 Asn His Leu Gln Leu Phe Pro Glu Leu Leu Phe Leu Gly Thr Ala Lys 115 120 125 Leu Tyr Arg Leu Asp Leu Ser Glu Asn Gln Ile Gln Ala Ile Pro Arg 130 135 140 Lys Ala Phe Arg Gly Ala Val Asp Ile Lys Asn Leu Gln Leu Asp Tyr 145 150 155 160 Asn Gln Ile Ser Cys Ile Glu Asp Gly Ala Phe Arg Ala Leu Arg Asp 165 170 175 Leu Glu Val Leu Thr Leu Asn Asn Asn Asn Ile Thr Arg Leu Ser Val 180 185 190 Ala Ser Phe Asn His Met Pro Lys Leu Arg Thr Phe Arg Leu His Ser 195 200 205 Asn Asn Leu Tyr Cys Asp Cys His Leu Ala Trp Leu Ser Asp Trp Leu 210 215 220 Arg Gln Arg Pro Arg Val Gly Leu Tyr Thr Gln Cys Met Gly Pro Ser 225 230 235 240 His Leu Arg Gly His Asn Val Ala Glu Val Gln Lys Arg Glu Phe Val 245 250 255 Cys Ser Gly His Gln Ser Phe Met Ala Pro Ser Cys Ser Val Leu His 260 265 270 Cys Pro Ala Ala Cys Thr Cys Ser Asn Asn Ile Val Asp Cys Arg Gly 275 280 285 Lys Gly Leu Thr Glu Ile Pro Thr Asn Leu Pro Glu Thr Ile Thr Glu 290 295 300 Ile Arg Leu Glu Gln Asn Thr Ile Lys Val Ile Pro Pro Gly Ala Phe 305 310 315 320 Ser Pro Tyr Lys Lys Leu Arg Arg Ile Asp Leu Ser Asn Asn Gln Ile 325 330 335 Ser Glu Leu Ala Pro Asp Ala Phe Gln Gly Leu Arg Ser Leu Asn Ser 340 345 350 Leu Val Leu Tyr Gly Asn Lys Ile Thr Glu Leu Pro Lys Ser Leu Phe 355 360 365 Glu Gly Leu Phe Ser Leu Gln Leu Leu Leu Leu Asn Ala Asn Lys Ile 370 375 380 Asn Cys Leu Arg Val Asp Ala Phe Gln Asp Leu His Asn Leu Asn Leu 385 390 395 400 Leu Ser Leu Tyr Asp Asn Lys Leu Gln Thr Ile Ala Lys Gly Thr Phe 405 410 415 Ser Pro Leu Arg Ala Ile Gln Thr Met His Leu Ala Gln Asn Pro Phe 420 425 430 Ile Cys Asp Cys His Leu Lys Trp Leu Ala Asp Tyr Leu His Thr Asn 435 440 445 Pro Ile Glu Thr Ser Gly Ala Arg Cys Thr Ser Pro Arg Arg Leu Ala 450 455 460 Asn Lys Arg Ile Gly Gln Ile Lys Ser Lys Lys Phe Arg Cys Ser Gly 465 470 475 480 Thr Glu Asp Tyr Arg Ser Lys Leu Ser Gly Asp Cys Phe Ala Asp Leu 485 490 495 Ala Cys Pro Glu Lys Cys Arg Cys Glu Gly Thr Thr Val Asp Cys Ser 500 505 510 Asn Gln Lys Leu Asn Lys Ile Pro Glu His Ile Pro Gln Tyr Thr Ala 515 520 525 Glu Leu Arg Leu Asn Asn Asn Glu Phe Thr Val Leu Glu Ala Thr Gly 530 535 540 Ile Phe Lys Lys Leu Pro Gln Leu Arg Lys Ile Asn Phe Ser Asn Asn 545 550 555 560 Lys Ile Thr Asp Ile Glu Glu Gly Ala Phe Glu Gly Ala Ser Gly Val 565 570 575 Asn Glu Ile Leu Leu Thr Ser Asn Arg Leu Glu Asn Val Gln His Lys 580 585 590 Met Phe Lys Gly Leu Glu Ser Leu Lys Thr Leu Met Leu Arg Ser Asn 595 600 605 Arg Ile Thr Cys Val Gly Asn Asp Ser Phe Ile Gly Leu Ser Ser Val 610 615 620 Arg Leu Leu Ser Leu Tyr Asp Asn Gln Ile Thr Thr Val Ala Pro Gly 625 630 635 640 Ala Phe Asp Thr Leu His Ser Leu Ser Thr Leu Asn Leu Leu Ala Asn 645 650 655 Pro Phe Asn Cys Asn Cys Tyr Leu Ala Trp Leu Gly Glu Trp Leu Arg 660 665 670 Lys Lys Arg Ile Val Thr Gly Asn Pro Arg Cys Gln Lys Pro Tyr Phe 675 680 685 Leu Lys Glu Ile Pro Ile Gln Asp Val Ala Ile Gln Asp Phe Thr Cys 690 695 700 Asp Asp Gly Asn Asp Asp Asn Ser Cys Ser Pro Leu Ser Arg Cys Pro 705 710 715 720 Thr Glu Cys Thr Cys Leu Asp Thr Val Val Arg Cys Ser Asn Lys Gly 725 730 735 Leu Lys Val Leu Pro Lys Gly Ile Pro Arg Asp Val Thr Glu Leu Tyr 740 745 750 Leu Asp Gly Asn Gln Phe Thr Leu Val Pro Lys Glu Leu Ser Asn Tyr 755 760 765 Lys His Leu Thr Leu Ile Asp Leu Ser Asn Asn Arg Ile Ser Thr Leu 770 775 780 Ser Asn Gln Ser Phe Ser Asn Met Thr Gln Leu Leu Thr Leu Ile Leu 785 790 795 800 Ser Tyr Asn Arg Leu Arg Cys Ile Pro Pro Arg Thr Phe Asp Gly Leu 805 810 815 Lys Ser Leu Arg Leu Leu Ser Leu His Gly Asn Asp Ile Ser Val Val 820 825 830 Pro Glu Gly Ala Phe Asn Asp Leu Ser Ala Leu Ser His Leu Ala Ile 835 840 845 Gly Ala Asn Pro Leu Tyr Cys Asp Cys Asn Met Gln Trp Leu Ser Asp 850 855 860 Trp Val Lys Ser Glu Tyr Lys Glu Pro Gly Ile Ala Arg Cys Ala Gly 865 870 875 880 Pro Gly Glu Met Ala Asp Lys Leu Leu Leu Thr Thr Pro Ser Lys Lys 885 890 895 Phe Thr Cys Gln Gly Pro Val Asp Val Asn Ile Leu Ala Lys Cys Asn 900 905 910 Pro Cys Leu Ser Asn Pro Cys Lys Asn Asp Gly Thr Cys Asn Ser Asp 915 920 925 Pro Val Asp Phe Tyr Arg Cys Thr Cys Pro Tyr Gly Phe Lys Gly Gln 930 935 940 Asp Cys Asp Val Pro Ile His Ala Cys Ile Ser Asn Pro Cys Lys His 945 950 955 960 Gly Gly Thr Cys His Leu Lys Glu Gly Glu Glu Asp Gly Phe Trp Cys 965 970 975 Ile Cys Ala Asp Gly Phe Glu Gly Glu Asn Cys Glu Val Asn Val Asp 980 985 990 Asp Cys Glu Asp Asn Asp Cys Glu Asn Asn Ser Thr Cys Val Asp Gly 995 1000 1005 Ile Asn Asn Tyr Thr Cys Leu Cys Pro Pro Glu Tyr Thr Gly Glu Leu 1010 1015 1020 Cys Glu Glu Lys Leu Asp Phe Cys Ala Gln Asp Leu Asn Pro Cys Gln 1025 1030 1035 1040 His Asp Ser Lys Cys Ile Leu Thr Pro Lys Gly Phe Lys Cys Asp Cys 1045 1050 1055 Thr Pro Gly Tyr Val Gly Glu His Cys Asp Ile Asp Phe Asp Asp Cys 1060 1065 1070 Gln Asp Asn Lys Cys Lys Asn Gly Ala His Cys Thr Asp Ala Val Asn 1075 1080 1085 Gly Tyr Thr Cys Ile Cys Pro Glu Gly Tyr Ser Gly Leu Phe Cys Glu 1090 1095 1100 Phe Ser Pro Pro Met Val Leu Pro Arg Thr Ser Pro Cys Asp Asn Phe 1105 1110 1115 1120 Asp Cys Gln Asn Gly Ala Gln Cys Ile Val Arg Ile Asn Glu Pro Ile 1125 1130 1135 Cys Gln Cys Leu Pro Gly Tyr Gln Gly Glu Lys Cys Glu Lys Leu Val 1140 1145 1150 Ser Val Asn Phe Ile Asn Lys Glu Ser Tyr Leu Gln Ile Ser Ser Ala 1155 1160 1165 Lys Val Arg Pro Gln Thr Asn Ile Thr Leu Gln Ile Ala Thr Asp Glu 1170 1175 1180 Asp Ser Gly Ile Leu Leu Tyr Lys Gly Asp Lys Asp His Ile Ala Val 1185 1190 1195 1200 Glu Leu Tyr Arg Gly Arg Val Val Ala Ser Tyr Asp Thr Gly Ser His 1205 1210 1215 Pro Ala Ser Ala Ile Tyr Ser Val Glu Thr Ile Asn Asp Gly Asn Phe 1220 1225 1230 His Ile Val Glu Leu Leu Ala Leu Asp Gln Ser Leu Ser Leu Ser Val 1235 1240 1245 Asp Gly Gly Asn Pro Lys Ile Ile Thr Asn Leu Ser Lys Gln Ser Thr 1250 1255 1260 Leu Asn Phe Asp Ser Pro Leu Tyr Val Gly Gly Met Pro Gly Lys Ser 1265 1270 1275 1280 Asn Val Ala Ser Leu Arg Gln Ala Pro Gly Gln Asn Gly Thr Ser Phe 1285 1290 1295 His Gly Cys Ile Arg Asn Leu Tyr Ile Asn Ser Glu Leu Gln Asp Phe 1300 1305 1310 Gln Lys Val Pro Met Gln Thr Gly Ile Leu Pro Gly Cys Glu Pro Cys 1315 1320 1325 His Lys Lys Val Cys Ala His Gly Thr Cys Gln Pro Ser Ser Gln Ala 1330 1335 1340 Gly Phe Thr Cys Glu Cys Gln Glu Gly Trp Met Gly Pro Leu Cys Asp 1345 1350 1355 1360 Gln Arg Thr Asn Asp Pro Cys Leu Gly Asn Lys Cys Val His Gly Thr 1365 1370 1375 Cys Leu Pro Ile Asn Ala Phe Ser Tyr Ser Cys Lys Cys Leu Glu Gly 1380 1385 1390 His Gly Gly Val Leu Cys Asp Glu Glu Glu Asp Leu Phe Asn Pro Cys 1395 1400 1405 Gln Ala Ile Lys Cys Lys His Gly Lys Cys Arg Leu Ser Gly Leu Gly 1410 1415 1420 Gln Pro Tyr Cys Glu Cys Ser Ser Gly Tyr Thr Gly Asp Ser Cys Asp 1425 1430 1435 1440 Arg Glu Ile Ser Cys Arg Gly Glu Arg Ile Arg Asp Tyr Tyr Gln Lys 1445 1450 1455 Gln Gln Gly Tyr Ala Ala Cys Gln Thr Thr Lys Lys Val Ser Arg Leu 1460 1465 1470 Glu Cys Arg Gly Gly Cys Ala Gly Gly Gln Cys Cys Gly Pro Leu Arg 1475 1480 1485 Ser Lys Arg Arg Lys Tyr Ser Phe Glu Cys Thr Asp Gly Ser Ser Phe 1490 1495 1500 Val Asp Glu Val Glu Lys Val Val Lys Cys Gly Cys Thr Arg Cys Val 1505 1510 1515 1520 Ser <210> 4 <211> 6398 <212> DNA Homo sapiens slit homolog 2 (Drosophila) (SLIT2), transcript variant 1, mRNA <400> 4 cggattcatc ctcaggacct aaagttgccc aaggagctcc tgctctgcca gaggagggtg 60 ggagggcgg tgggaggcgt gtgcctgagt gggctctact gccttgttcc atattatttg 120 gtgcacattt tccctggcac tctgggttgc tagccccgcc gggcactggg cctcagacac 180 tgcgcggttc cctcggagca gcaagctaaa gaaagccccc agtgccggcg aggaaggagg 240 cggcggggaa agatgcgcgg cgttggctgg cagatgctgt ccctgtcgct ggggttagtg 300 ctggcgatcc tgaacaaggt ggcaccgcag gcgtgcccgg cgcagtgctc ttgctcgggc 360 agcacagtgg actgtcacgg gctggcgctg cgcagcgtgc ccaggaatat cccccgcaac 420 accgagagac tggatttaaa tggaaataac atcacaagaa ttacgaagac agattttgct 480 ggtcttagac atctaagagt tcttcagctt atggagaata agattagcac cattgaaaga 540 ggagcattcc aggatcttaa agaactagag agactgcgtt taaacagaaa tcaccttcag 600 ctgtttcctg agttgctgtt tcttgggact gcgaagctat acaggcttga tctcagtgaa 660 aaccaaattc aggcaatccc aaggaaagct ttccgtgggg cagttgacat aaaaaatttg 720 caactggatt acaaccagat cagctgtatt gaagatgggg cattcagggc tctccgggac 780 ctggaagtgc tcactctcaa caataacaac attactagac tttctgtggc aagtttcaac 840 catatgccta aacttaggac ttttcgactg cattcaaaca acctgtattg tgactgccac 900 ctggcctggc tctccgactg gcttcgccaa aggcctcggg ttggtctgta cactcagtgt 960 atgggcccct cccacctgag aggccataat gtagccgagg ttcaaaaacg agaatttgtc 1020 tgcagtggtc accagtcatt tatggctcct tcttgtagtg ttttgcactg ccctgccgcc 1080 tgtacctgta gcaacaatat cgtagactgt cgtgggaaag gtctcactga gatccccaca 1140 aatcttccag agaccatcac agaaatacgt ttggaacaga acacaatcaa agtcatccct 1200 cctggagctt tctcaccata taaaaagctt agacgaattg acctgagcaa taatcagatc 1260 tctgaacttg caccagatgc tttccaagga ctacgctctc tgaattcact tgtcctctat 1320 ggaaataaaa tcacagaact ccccaaaagt ttatttgaag gactgttttc cttacagctc 1380 ctattattga atgccaacaa gataaactgc cttcgggtag atgcttttca ggatctccac 1440 aacttgaacc ttctctccct atatgacaac aagcttcaga ccatcgccaa ggggaccttt 1500 tcacctcttc gggccattca aactatgcat ttggcccaga acccctttat ttgtgactgc 1560 catctcaagt ggctagcgga ttatctccat accaacccga ttgagaccag tggtgcccgt 1620 tgcaccagcc cccgccgcct ggcaaacaaa agaattggac agatcaaaag caagaaattc 1680 cgttgttcag ctaaagaaca gtatttcatt ccaggtacag aagattatcg atcaaaatta 1740 agtggagact gctttgcgga tctggcttgc cctgaaaagt gtcgctgtga aggaaccaca 1800 gtagattgct ctaatcaaaa gctcaacaaa atcccggagc acattcccca gtacactgca 1860 gagttgcgtc tcaataataa tgaatttacc gtgttggaag ccacaggaat ctttaagaaa 1920 cttcctcaat tacgtaaaat aaactttagc aacaataaga tcacagatat tgaggaggga 1980 gcatttgaag gagcatctgg tgtaaatgaa atacttctta cgagtaatcg tttggaaaat 2040 gtgcagcata agatgttcaa gggattggaa agcctcaaaa ctttgatgtt gagaagcaat 2100 cgaataacct gtgtggggaa tgacagtttc ataggactca gttctgtgcg tttgctttct 2160 ttgtatgata atcaaattac tacagttgca ccaggggcat ttgatactct ccattcttta 2220 tctactctaa acctcttggc caatcctttt aactgtaact gctacctggc ttggttggga 2280 gagtggctga gaaagaagag aattgtcacg ggaaatccta gatgtcaaaa accatacttc 2340 ctgaaagaaa tacccatcca ggatgtggcc attcaggact tcacttgtga tgacggaaat 2400 gatgacaata gttgctcccc actttctcgc tgtcctactg aatgtacttg cttggataca 2460 gtcgtccgat gtagcaacaa gggtttgaag gtcttgccga aaggtattcc aagagatgtc 2520 acagagttgt atctggatgg aaaccaattt acactggttc ccaaggaact ctccaactac 2580 aaacatttaa cacttataga cttaagtaac aacagaataa gcacgctttc taatcagagc 2640 ttcagcaaca tgacccagct cctcacctta attcttagtt acaaccgtct gagatgtatt 2700 cctcctcgca cctttgatgg attaaagtct cttcgattac tttctctaca tggaaatgac 2760 atttctgttg tgcctgaagg tgctttcaat gatctttctg cattatcaca tctagcaatt 2820 ggagccaacc ctctttactg tgattgtaac atgcagtggt tatccgactg ggtgaagtcg 2880 gaatataagg agcctggaat tgctcgttgt gctggtcctg gagaaatggc agataaactt 2940 ttactcacaa ctccctccaa aaaatttacc tgtcaaggtc ctgtggatgt caatattcta 3000 gctaagtgta acccctgcct atcaaatccg tgtaaaaatg atggcacatg taatagtgat 3060 ccagttgact tttaccgatg cacctgtcca tatggtttca aggggcagga ctgtgatgtc 3120 ccaattcatg cctgcatcag taacccatgt aaacatggag gaacttgcca cttaaaggaa 3180 ggagaagaag atggattctg gtgtatttgt gctgatggat ttgaaggaga aaattgtgaa 3240 gtcaacgttg atgattgtga agataatgac tgtgaaaata attctacatg tgtcgatggc 3300 attaataact acacatgcct ttgcccacct gagtatacag gtgagttgtg tgaggagaag 3360 ctggacttct gtgcccagga cctgaacccc tgccagcacg attcaaagtg catcctaact 3420 ccaaagggat tcaaatgtga ctgcacacca gggtacgtag gtgaacactg cgacatcgat 3480 tttgacgact gccaagacaa caagtgtaaa aacggagccc actgcacaga tgcagtgaac 3540 ggctatacgt gcatatgccc cgaaggttac agtggcttgt tctgtgagtt ttctccaccc 3600 atggtcctcc ctcgtaccag cccctgtgat aattttgatt gtcagaatgg agctcagtgt 3660 atcgtcagaa taaatgagcc aatatgtcag tgtttgcctg gctatcaggg agaaaagtgt 3720 gaaaaattgg ttagtgtgaa ttttataaac aaagagtctt atcttcagat tccttcagcc 3780 aaggttcggc ctcagacgaa cataacactt cagattgcca cagatgaaga cagcggaatc 3840 ctcctgtata agggtgacaa agaccatatc gcggtagaac tctatcgggg gcgtgttcgt 3900 gccagctatg acaccggctc tcatccagct tctgccattt acagtgtgga gacaatcaat 3960 gatggaaact tccacattgt ggaactactt gccttggatc agagtctctc tttgtccgtg 4020 gatggtggga accccaaaat catcactaac ttgtcaaagc agtccactct gaattttgac 4080 tctccactct atgtaggagg catgccaggg aagagtaacg tggcatctct gcgccaggcc 4140 cctgggcaga acggaaccag cttccacggc tgcatccgga acctttacat caacagtgag 4200 ctgcaggact tccagaaggt gccgatgcaa acaggcattt tgcctggctg tgagccatgc 4260 cacaagaagg tgtgtgccca tggcacatgc cagcccagca gccaggcagg cttcacctgc 4320 gagtgccagg aaggatggat ggggcccctc tgtgaccaac ggaccaatga cccttgcctt 4380 ggaaataaat gcgtacatgg cacctgcttg cccatcaatg cgttctccta cagctgtaag 4440 tgcttggagg gccatggagg tgtcctctgt gatgaagagg aggatctgtt taacccatgc 4500 caggcgatca agtgcaagca tgggaagtgc aggctttcag gtctggggca gccctactgt 4560 gaatgcagca gtggatacac gggggacagc tgtgatcgag aaatctcttg tcgaggggaa 4620 aggataagag attattacca aaagcagcag ggctatgctg cttgccaaac aaccaagaag 4680 gtgtcccgat tagagtgcag aggtgggtgt gcaggagggc agtgctgtgg accgctgagg 4740 agcaagcggc ggaaatactc tttcgaatgc actgacggct cctcctttgt ggacgaggtt 4800 gagaaagtgg tgaagtgcgg ctgtacgagg tgtgtgtcct aaacacactc ccggcagctc 4860 tgtctttgga aaaggttgta tacttcttga ccgtgtggga ctaatgaatg cttcatagtg 4920 gaaatatttg aaatatattg taaaatacag aacagactta tttttattat gagaataaag 4980 actttttttc tgcatttgga aaaaaaaaaa aagaaatgct tgaactaaag cttcccctat 5040 gctggagaag tatgaagaaa gatatacctg gagacattag aacagcgatg ggaaccattg 5100 caactcgggt ccatctttgt aacatgctga agacaagcag aagcacatgc acgagggaca 5160 gaggagctac tgtgcactgc tgtgaaattg cccagagcat aaaacctgtg taccctcctt 5220 cacatcaacc aagttcacta ggacatacca agcacatgcg tgtgaatgag gatgcaaggc 5280 aaggaatgg aactccagaa ttcacagata gaacaaatgg atgagaaata tttcatgcaa 5340 aatataaagt gtcctgagga tctgggttcc atttacgaga ggaaatgaaa gtgctaaaat 5400 aaattttatc ttccttttaa atgtcagcat gtcagcagaa gcagcacaca aaagtcttta 5460 ccattttcca gtattaattt ttttgtaata taaatgataa aggagatgat aaagaaccaa 5520 tagattattg ataaataaat tagtaataat atgatttttt gtttctatga gttctaaaca 5580 gccctataca gtattcagtt tccatgagaa atatttattg tatcaaagta cattgtactt 5640 aacattttag gccatccttt tactgttcct tgatgcttta atatatatta atttataatt 5700 atcctgatat ttttgtacat ttttcaaact taaaaatcag gatttttttt tttttttttt 5760 tggcaatagt actaacatag ggttttatct tgggatacat atgtattggt ctgtttgttg 5820 accttgacat ctgatcactg atgtcaatga cctactggcc tcattcagga cacctgcaga 5880 gagtatgcaa agtccgagag aggaaaacag aaatctgtat gtgagatggt cattgtacag 5940 aaagaagtgg cccctctgca acatgtcctc acagaaacga aatggtgtgt agcaatcaac 6000 actagaaagt agaccttttg caaattaata tgtccttgac cttttttgcc cttttgtggg 6060 ggtgaggtgg ggataaaaag actgtcatat caagaactgt gacttttctt tccctcaaac 6120 aataaaactc ctttattatc ttaatgctcc catgttaaca tgtttgctgc taaattacaa 6180 tgtagaattg ataatggttt atagtgaact gtgctcttcc ctcattaaaa tcccagggtg 6240 ccctgtaaag atgcagatgt ttcttcctga aaacttcttt ttttacaaag aaaattagat 6300 gtacatgtat aattcagtgt gctttgtctt tctccagatt aatatcggtt acactgctga 6360 tgtttgtaaa ttaaacagat atttacttca ttaaaaaa 6398 <210> 5 <211> 6386 <212> DNA Homo sapiens slit homolog 2 (Drosophila) (SLIT2), transcript variant 2, mRNA <400> 5 cggattcatc ctcaggacct aaagttgccc aaggagctcc tgctctgcca gaggagggtg 60 ggagggcgg tgggaggcgt gtgcctgagt gggctctact gccttgttcc atattatttg 120 gtgcacattt tccctggcac tctgggttgc tagccccgcc gggcactggg cctcagacac 180 tgcgcggttc cctcggagca gcaagctaaa gaaagccccc agtgccggcg aggaaggagg 240 cggcggggaa agatgcgcgg cgttggctgg cagatgctgt ccctgtcgct ggggttagtg 300 ctggcgatcc tgaacaaggt ggcaccgcag gcgtgcccgg cgcagtgctc ttgctcgggc 360 agcacagtgg actgtcacgg gctggcgctg cgcagcgtgc ccaggaatat cccccgcaac 420 accgagagac tggatttaaa tggaaataac atcacaagaa ttacgaagac agattttgct 480 ggtcttagac atctaagagt tcttcagctt atggagaata agattagcac cattgaaaga 540 ggagcattcc aggatcttaa agaactagag agactgcgtt taaacagaaa tcaccttcag 600 ctgtttcctg agttgctgtt tcttgggact gcgaagctat acaggcttga tctcagtgaa 660 aaccaaattc aggcaatccc aaggaaagct ttccgtgggg cagttgacat aaaaaatttg 720 caactggatt acaaccagat cagctgtatt gaagatgggg cattcagggc tctccgggac 780 ctggaagtgc tcactctcaa caataacaac attactagac tttctgtggc aagtttcaac 840 catatgccta aacttaggac ttttcgactg cattcaaaca acctgtattg tgactgccac 900 ctggcctggc tctccgactg gcttcgccaa aggcctcggg ttggtctgta cactcagtgt 960 atgggcccct cccacctgag aggccataat gtagccgagg ttcaaaaacg agaatttgtc 1020 tgcagtgatg aggaagaagg tcaccagtca tttatggctc cttcttgtag tgttttgcac 1080 tgccctgccg cctgtacctg tagcaacaat atcgtagact gtcgtgggaa aggtctcact 1140 gagatcccca caaatcttcc agagaccatc acagaaatac gtttggaaca gaacacaatc 1200 aaagtcatcc ctcctggagc tttctcacca tataaaaagc ttagacgaat tgacctgagc 1260 aataatcaga tctctgaact tgcaccagat gctttccaag gactacgctc tctgaattca 1320 cttgtcctct atggaaataa aatcacagaa ctccccaaaa gtttatttga aggactgttt 1380 tccttacagc tcctattatt gaatgccaac aagataaact gccttcgggt agatgctttt 1440 caggatctcc acaacttgaa ccttctctcc ctatatgaca acaagcttca gaccatcgcc 1500 aaggggacct tttcacctct tcgggccatt caaactatgc atttggccca gaaccccttt 1560 atttgtgact gccatctcaa gtggctagcg gattatctcc ataccaaccc gattgagacc 1620 agtggtgccc gttgcaccag cccccgccgc ctggcaaaca aaagaattgg acagatcaaa 1680 agcaagaaat tccgttgttc aggtacagaa gattatcgat caaaattaag tggagactgc 1740 tttgcggatc tggcttgccc tgaaaagtgt cgctgtgaag gaaccacagt agattgctct 1800 aatcaaaagc tcaacaaaat cccggagcac attccccagt acactgcaga gttgcgtctc 1860 aataataatg aatttaccgt gttggaagcc acaggaatct ttaagaaact tcctcaatta 1920 cgtaaaataa actttagcaa caataagatc acagatattg aggagggagc atttgaagga 1980 gcatctggtg taaatgaaat acttcttacg agtaatcgtt tggaaaatgt gcagcataag 2040 atgttcaagg gattggaaag cctcaaaact ttgatgttga gaagcaatcg aataacctgt 2100 gtggggaatg acagtttcat aggactcagt tctgtgcgtt tgctttcttt gtatgataat 2160 caaattacta cagttgcacc aggggcattt gatactctcc attctttatc tactctaaac 2220 ctcttggcca atccttttaa ctgtaactgc tacctggctt ggttgggaga gtggctgaga 2280 aagaagagaa ttgtcacggg aaatcctaga tgtcaaaaac catacttcct gaaagaaata 2340 cccatccagg atgtggccat tcaggacttc acttgtgatg acggaaatga tgacaatagt 2400 tgctccccac tttctcgctg tcctactgaa tgtacttgct tggatacagt cgtccgatgt 2460 agcaacaagg gtttgaaggt cttgccgaaa ggtattccaa gagatgtcac agagttgtat 2520 ctggatggaa accaatttac actggttccc aaggaactct ccaactacaa acatttaaca 2580 cttatagact taagtaacaa cagaataagc acgctttcta atcagagctt cagcaacatg 2640 acccagctcc tcaccttaat tcttagttac aaccgtctga gatgtattcc tcctcgcacc 2700 tttgatggat taaagtctct tcgattactt tctctacatg gaaatgacat ttctgttgtg 2760 cctgaaggtg ctttcaatga tctttctgca ttatcacatc tagcaattgg agccaaccct 2820 ctttactgtg attgtaacat gcagtggtta tccgactggg tgaagtcgga atataaggag 2880 cctggaattg ctcgttgtgc tggtcctgga gaaatggcag ataaactttt actcacaact 2940 ccctccaaaa aatttacctg tcaaggtcct gtggatgtca atattctagc taagtgtaac 3000 ccctgcctat caaatccgtg taaaaatgat ggcacatgta atagtgatcc agttgacttt 3060 taccgatgca cctgtccata tggtttcaag gggcaggact gtgatgtccc aattcatgcc 3120 tgcatcagta acccatgtaa acatggagga acttgccact taaaggaagg agaagaagat 3180 ggattctggt gtatttgtgc tgatggattt gaaggagaaa attgtgaagt caacgttgat 3240 gattgtgaag ataatgactg tgaaaataat tctacatgtg tcgatggcat taataactac 3300 acatgccttt gcccacctga gtatacaggt gagttgtgtg aggagaagct ggacttctgt 3360 gcccaggacc tgaacccctg ccagcacgat tcaaagtgca tcctaactcc aaagggattc 3420 aaatgtgact gcacaccagg gtacgtaggt gaacactgcg acatcgattt tgacgactgc 3480 caagacaaca agtgtaaaaa cggagcccac tgcacagatg cagtgaacgg ctatacgtgc 3540 atatgccccg aaggttacag tggcttgttc tgtgagtttt ctccacccat ggtcctccct 3600 cgtaccagcc cctgtgataa ttttgattgt cagaatggag ctcagtgtat cgtcagaata 3660 aatgagccaa tatgtcagtg tttgcctggc tatcagggag aaaagtgtga aaaattggtt 3720 agtgtgaatt ttataaacaa agagtcttat cttcagattc cttcagccaa ggttcggcct 3780 cagacgaaca taacacttca gattgccaca gatgaagaca gcggaatcct cctgtataag 3840 ggtgacaaag accatatcgc ggtagaactc tatcgggggc gtgttcgtgc cagctatgac 3900 accggctctc atccagcttc tgccatttac agtgtggaga caatcaatga tggaaacttc 3960 cacattgtgg aactacttgc cttggatcag agtctctctt tgtccgtgga tggtgggaac 4020 cccaaaatca tcactaactt gtcaaagcag tccactctga attttgactc tccactctat 4080 gtaggaggca tgccagggaa gagtaacgtg gcatctctgc gccaggcccc tgggcagaac 4140 ggaaccagct tccacggctg catccggaac ctttacatca acagtgagct gcaggacttc 4200 cagaaggtgc cgatgcaaac aggcattttg cctggctgtg agccatgcca caagaaggtg 4260 tgtgcccatg gcacatgcca gcccagcagc caggcaggct tcacctgcga gtgccaggaa 4320 ggatggatgg ggcccctctg tgaccaacgg accaatgacc cttgccttgg aaataaatgc 4380 gtacatggca cctgcttgcc catcaatgcg ttctcctaca gctgtaagtg cttggagggc 4440 catggaggtg tcctctgtga tgaagaggag gatctgttta acccatgcca ggcgatcaag 4500 tgcaagcatg ggaagtgcag gctttcaggt ctggggcagc cctactgtga atgcagcagt 4560 ggatacacgg gggacagctg tgatcgagaa atctcttgtc gaggggaaag gataagagat 4620 tattaccaaa agcagcaggg ctatgctgct tgccaaacaa ccaagaaggt gtcccgatta 4680 gagtgcagag gtgggtgtgc aggagggcag tgctgtggac cgctgaggag caagcggcgg 4740 aaatactctt tcgaatgcac tgacggctcc tcctttgtgg acgaggttga gaaagtggtg 4800 aagtgcggct gtacgaggtg tgtgtcctaa acacactccc ggcagctctg tctttggaaa 4860 aggttgtata cttcttgacc gtgtgggact aatgaatgct tcatagtgga aatatttgaa 4920 atatattgta aaatacagaa cagacttatt tttattatga gaataaagac tttttttctg 4980 catttggaaa aaaaaaaaaa gaaatgcttg aactaaagct tcccctatgc tggagaagta 5040 tgaagaaaga tatacctgga gacattagaa cagcgatggg aaccattgca actcgggtcc 5100 atctttgtaa catgctgaag acaagcagaa gcacatgcac gagggacaga ggagctactg 5160 tgcactgctg tgaaattgcc cagagcataa aacctgtgta ccctccttca catcaaccaa 5220 gttcactagg acataccaag cacatgcgtg tgaatgagga tgcaaggcaa gagaatggaa 5280 ctccagaatt cacagataga acaaatggat gagaaatatt tcatgcaaaa tataaagtgt 5340 cctgaggatc tgggttccat ttacgagagg aaatgaaagt gctaaaataa attttatctt 5400 ccttttaaat gtcagcatgt cagcagaagc agcacacaaa agtctttacc attttccagt 5460 attaattttt ttgtaatata aatgataaag gagatgataa agaaccaata gattattgat 5520 aaataaatta gtaataatat gattttttgt ttctatgagt tctaaacagc cctatacagt 5580 attcagtttc catgagaaat atttattgta tcaaagtaca ttgtacttaa cattttaggc 5640 catcctttta ctgttccttg atgctttaat atatattaat ttataattat cctgatattt 5700 ttgtacattt ttcaaactta aaaatcagga tttttttttt tttttttttg gcaatagtac 5760 taacataggg ttttatcttg ggatacatat gtattggtct gtttgttgac cttgacatct 5820 gatcactgat gtcaatgacc tactggcctc attcaggaca cctgcagaga gtatgcaaag 5880 tccgagagag gaaaacagaa atctgtatgt gagatggtca ttgtacagaa agaagtggcc 5940 cctctgcaac atgtcctcac agaaacgaaa tggtgtgtag caatcaacac tagaaagtag 6000 accttttgca aattaatatg tccttgacct tttttgccct tttgtggggg tgaggtgggg 6060 ataaaaagac tgtcatatca agaactgtga cttttctttc cctcaaacaa taaaactcct 6120 ttattatctt aatgctccca tgttaacatg tttgctgcta aattacaatg tagaattgat 6180 aatggtttat agtgaactgt gctcttccct cattaaaatc ccagggtgcc ctgtaaagat 6240 gcagatgttt cttcctgaaa acttcttttt ttacaaagaa aattagatgt acatgtataa 6300 ttcagtgtgc tttgtctttc tccagattaa tatcggttac actgctgatg tttgtaaatt 6360 aaacagatat ttacttcatt aaaaaa 6386 <210> 6 <211> 6374 <212> DNA Homo sapiens slit homolog 2 (Drosophila) (SLIT2), transcript variant 3, mRNA <400> 6 cggattcatc ctcaggacct aaagttgccc aaggagctcc tgctctgcca gaggagggtg 60 ggagggcgg tgggaggcgt gtgcctgagt gggctctact gccttgttcc atattatttg 120 gtgcacattt tccctggcac tctgggttgc tagccccgcc gggcactggg cctcagacac 180 tgcgcggttc cctcggagca gcaagctaaa gaaagccccc agtgccggcg aggaaggagg 240 cggcggggaa agatgcgcgg cgttggctgg cagatgctgt ccctgtcgct ggggttagtg 300 ctggcgatcc tgaacaaggt ggcaccgcag gcgtgcccgg cgcagtgctc ttgctcgggc 360 agcacagtgg actgtcacgg gctggcgctg cgcagcgtgc ccaggaatat cccccgcaac 420 accgagagac tggatttaaa tggaaataac atcacaagaa ttacgaagac agattttgct 480 ggtcttagac atctaagagt tcttcagctt atggagaata agattagcac cattgaaaga 540 ggagcattcc aggatcttaa agaactagag agactgcgtt taaacagaaa tcaccttcag 600 ctgtttcctg agttgctgtt tcttgggact gcgaagctat acaggcttga tctcagtgaa 660 aaccaaattc aggcaatccc aaggaaagct ttccgtgggg cagttgacat aaaaaatttg 720 caactggatt acaaccagat cagctgtatt gaagatgggg cattcagggc tctccgggac 780 ctggaagtgc tcactctcaa caataacaac attactagac tttctgtggc aagtttcaac 840 catatgccta aacttaggac ttttcgactg cattcaaaca acctgtattg tgactgccac 900 ctggcctggc tctccgactg gcttcgccaa aggcctcggg ttggtctgta cactcagtgt 960 atgggcccct cccacctgag aggccataat gtagccgagg ttcaaaaacg agaatttgtc 1020 tgcagtggtc accagtcatt tatggctcct tcttgtagtg ttttgcactg ccctgccgcc 1080 tgtacctgta gcaacaatat cgtagactgt cgtgggaaag gtctcactga gatccccaca 1140 aatcttccag agaccatcac agaaatacgt ttggaacaga acacaatcaa agtcatccct 1200 cctggagctt tctcaccata taaaaagctt agacgaattg acctgagcaa taatcagatc 1260 tctgaacttg caccagatgc tttccaagga ctacgctctc tgaattcact tgtcctctat 1320 ggaaataaaa tcacagaact ccccaaaagt ttatttgaag gactgttttc cttacagctc 1380 ctattattga atgccaacaa gataaactgc cttcgggtag atgcttttca ggatctccac 1440 aacttgaacc ttctctccct atatgacaac aagcttcaga ccatcgccaa ggggaccttt 1500 tcacctcttc gggccattca aactatgcat ttggcccaga acccctttat ttgtgactgc 1560 catctcaagt ggctagcgga ttatctccat accaacccga ttgagaccag tggtgcccgt 1620 tgcaccagcc cccgccgcct ggcaaacaaa agaattggac agatcaaaag caagaaattc 1680 cgttgttcag gtacagaaga ttatcgatca aaattaagtg gagactgctt tgcggatctg 1740 gcttgccctg aaaagtgtcg ctgtgaagga accacagtag attgctctaa tcaaaagctc 1800 aacaaaatcc cggagcacat tccccagtac actgcagagt tgcgtctcaa taataatgaa 1860 tttaccgtgt tggaagccac aggaatcttt aagaaacttc ctcaattacg taaaataaac 1920 tttagcaaca ataagatcac agatattgag gagggagcat ttgaaggagc atctggtgta 1980 aatgaaatac ttcttacgag taatcgtttg gaaaatgtgc agcataagat gttcaaggga 2040 ttggaaagcc tcaaaacttt gatgttgaga agcaatcgaa taacctgtgt ggggaatgac 2100 agtttcatag gactcagttc tgtgcgtttg ctttctttgt atgataatca aattactaca 2160 gttgcaccag gggcatttga tactctccat tctttatcta ctctaaacct cttggccaat 2220 ccttttaact gtaactgcta cctggcttgg ttgggagagt ggctgagaaa gaagagaatt 2280 gtcacgggaa atcctagatg tcaaaaacca tacttcctga aagaaatacc catccaggat 2340 gtggccattc aggacttcac ttgtgatgac ggaaatgatg acaatagttg ctccccactt 2400 tctcgctgtc ctactgaatg tacttgcttg gatacagtcg tccgatgtag caacaagggt 2460 ttgaaggtct tgccgaaagg tattccaaga gatgtcacag agttgtatct ggatggaaac 2520 caatttacac tggttcccaa ggaactctcc aactacaaac atttaacact tatagactta 2580 agtaacaaca gaataagcac gctttctaat cagagcttca gcaacatgac ccagctcctc 2640 accttaattc ttagttacaa ccgtctgaga tgtattcctc ctcgcacctt tgatggatta 2700 aagtctcttc gattactttc tctacatgga aatgacattt ctgttgtgcc tgaaggtgct 2760 ttcaatgatc tttctgcatt atcacatcta gcaattggag ccaaccctct ttactgtgat 2820 tgtaacatgc agtggttatc cgactgggtg aagtcggaat ataaggagcc tggaattgct 2880 cgttgtgctg gtcctggaga aatggcagat aaacttttac tcacaactcc ctccaaaaaa 2940 tttacctgtc aaggtcctgt ggatgtcaat attctagcta agtgtaaccc ctgcctatca 3000 aatccgtgta aaaatgatgg cacatgtaat agtgatccag ttgactttta ccgatgcacc 3060 tgtccatatg gtttcaaggg gcaggactgt gatgtcccaa ttcatgcctg catcagtaac 3120 ccatgtaaac atggaggaac ttgccactta aaggaaggag aagaagatgg attctggtgt 3180 atttgtgctg atggatttga aggagaaaat tgtgaagtca acgttgatga ttgtgaagat 3240 aatgactgtg aaaataattc tacatgtgtc gatggcatta ataactacac atgcctttgc 3300 ccacctgagt atacaggtga gttgtgtgag gagaagctgg acttctgtgc ccaggacctg 3360 aacccctgcc agcacgattc aaagtgcatc ctaactccaa agggattcaa atgtgactgc 3420 acaccagggt acgtaggtga acactgcgac atcgattttg acgactgcca agacaacaag 3480 tgtaaaaacg gagcccactg cacagatgca gtgaacggct atacgtgcat atgccccgaa 3540 ggttacagtg gcttgttctg tgagttttct ccacccatgg tcctccctcg taccagcccc 3600 tgtgataatt ttgattgtca gaatggagct cagtgtatcg tcagaataaa tgagccaata 3660 tgtcagtgtt tgcctggcta tcagggagaa aagtgtgaaa aattggttag tgtgaatttt 3720 ataaacaaag agtcttatct tcagattcct tcagccaagg ttcggcctca gacgaacata 3780 acacttcaga ttgccacaga tgaagacagc ggaatcctcc tgtataaggg tgacaaagac 3840 catatcgcgg tagaactcta tcgggggcgt gttcgtgcca gctatgacac cggctctcat 3900 ccagcttctg ccatttacag tgtggagaca atcaatgatg gaaacttcca cattgtggaa 3960 ctacttgcct tggatcagag tctctctttg tccgtggatg gtgggaaccc caaaatcatc 4020 actaacttgt caaagcagtc cactctgaat tttgactctc cactctatgt aggaggcatg 4080 ccagggaaga gtaacgtggc atctctgcgc caggcccctg ggcagaacgg aaccagcttc 4140 cacggctgca tccggaacct ttacatcaac agtgagctgc aggacttcca gaaggtgccg 4200 atgcaaacag gcattttgcc tggctgtgag ccatgccaca agaaggtgtg tgcccatggc 4260 acatgccagc ccagcagcca ggcaggcttc acctgcgagt gccaggaagg atggatgggg 4320 cccctctgtg accaacggac caatgaccct tgccttggaa ataaatgcgt acatggcacc 4380 tgcttgccca tcaatgcgtt ctcctacagc tgtaagtgct tggagggcca tggaggtgtc 4440 ctctgtgatg aagaggagga tctgtttaac ccatgccagg cgatcaagtg caagcatggg 4500 aagtgcaggc tttcaggtct ggggcagccc tactgtgaat gcagcagtgg atacacgggg 4560 gacagctgtg atcgagaaat ctcttgtcga ggggaaagga taagagatta ttaccaaaag 4620 cagcagggct atgctgcttg ccaaacaacc aagaaggtgt cccgattaga gtgcagaggt 4680 gggtgtgcag gagggcagtg ctgtggaccg ctgaggagca agcggcggaa atactctttc 4740 gaatgcactg acggctcctc ctttgtggac gaggttgaga aagtggtgaa gtgcggctgt 4800 acgaggtgtg tgtcctaaac acactcccgg cagctctgtc tttggaaaag gttgtatact 4860 tcttgaccgt gtgggactaa tgaatgcttc atagtggaaa tatttgaaat atattgtaaa 4920 atacagaaca gacttatttt tattatgaga ataaagactt tttttctgca tttggaaaaa 4980 aaaaaaaaga aatgcttgaa ctaaagcttc ccctatgctg gagaagtatg aagaaagata 5040 tacctggaga cattagaaca gcgatgggaa ccattgcaac tcgggtccat ctttgtaaca 5100 tgctgaagac aagcagaagc acatgcacga gggacagagg agctactgtg cactgctgtg 5160 aaattgccca gagcataaaa cctgtgtacc ctccttcaca tcaaccaagt tcactaggac 5220 ataccaagca catgcgtgtg aatgaggatg caaggcaaga gaatggaact ccagaattca 5280 cagatagaac aaatggatga gaaatatttc atgcaaaata taaagtgtcc tgaggatctg 5340 ggttccattt acgagaggaa atgaaagtgc taaaataaat tttatcttcc ttttaaatgt 5400 cagcatgtca gcagaagcag cacacaaaag tctttaccat tttccagtat taattttttt 5460 gtaatataaa tgataaagga gatgataaag aaccaataga ttattgataa ataaattagt 5520 aataatatga ttttttgttt ctatgagttc taaacagccc tatacagtat tcagtttcca 5580 tgagaaatat ttattgtatc aaagtacatt gtacttaaca ttttaggcca tccttttact 5640 gttccttgat gctttaatat atattaattt ataattatcc tgatattttt gtacattttt 5700 caaacttaaa aatcaggatt tttttttttt tttttttggc aatagtacta acatagggtt 5760 ttatcttggg atacatatgt attggtctgt ttgttgacct tgacatctga tcactgatgt 5820 caatgaccta ctggcctcat tcaggacacc tgcagagagt atgcaaagtc cgagagagga 5880 aaacagaaat ctgtatgtga gatggtcatt gtacagaaag aagtggcccc tctgcaacat 5940 gtcctcacag aaacgaaatg gtgtgtagca atcaacacta gaaagtagac cttttgcaaa 6000 ttaatatgtc cttgaccttt tttgcccttt tgtgggggtg aggtggggat aaaaagactg 6060 tcatatcaag aactgtgact tttctttccc tcaaacaata aaactccttt attatcttaa 6120 tgctcccatg ttaacatgtt tgctgctaaa ttacaatgta gaattgataa tggtttatag 6180 tgaactgtgc tcttccctca ttaaaatccc agggtgccct gtaaagatgc agatgtttct 6240 tcctgaaaac ttcttttttt acaaagaaaa ttagatgtac atgtataatt cagtgtgctt 6300 tgtctttctc cagattaata tcggttacac tgctgatgtt tgtaaattaa acagatattt 6360 acttcattaa aaaa 6374 <210> 7 <211> 1651 <212> PRT <213> roundabout homolog 1 isoform a precursor [Homo sapiens] <400> 7 Met Lys Trp Lys His Val Pro Phe Leu Val Met Ile Ser Leu Leu Ser 1 5 10 15 Leu Ser Pro Asn His Leu Phe Leu Ala Gln Leu Ile Pro Asp Pro Glu 20 25 30 Asp Val Glu Arg Gly Asn Asp His Gly Thr Pro Ile Pro Thr Ser Asp 35 40 45 Asn Asp Asp Asn Ser Leu Gly Tyr Thr Gly Ser Arg Leu Arg Gln Glu 50 55 60 Asp Phe Pro Pro Arg Ile Val Glu His Pro Ser Asp Leu Ile Val Ser 65 70 75 80 Lys Gly Glu Pro Ala Thr Leu Asn Cys Lys Ala Glu Gly Arg Pro Thr 85 90 95 Pro Thr Ile Glu Trp Tyr Lys Gly Gly Glu Arg Val Glu Thr Asp Lys 100 105 110 Asp Asp Pro Arg Ser His Arg Met Leu Leu Pro Ser Gly Ser Leu Phe 115 120 125 Phe Leu Arg Ile Val His Gly Arg Lys Ser Arg Pro Asp Glu Gly Val 130 135 140 Tyr Val Cys Val Ala Arg Asn Tyr Leu Gly Glu Ala Val Ser His Asn 145 150 155 160 Ala Ser Leu Glu Val Ala Ile Leu Arg Asp Asp Phe Arg Gln Asn Pro 165 170 175 Ser Asp Val Met Val Ala Val Gly Glu Pro Ala Val Met Glu Cys Gln 180 185 190 Pro Pro Arg Gly His Pro Glu Pro Thr Ile Ser Trp Lys Lys Asp Gly 195 200 205 Ser Pro Leu Asp Asp Lys Asp Glu Arg Ile Thr Ile Arg Gly Gly Lys 210 215 220 Leu Met Ile Thr Tyr Thr Arg Lys Ser Asp Ala Gly Lys Tyr Val Cys 225 230 235 240 Val Gly Thr Asn Met Val Gly Glu Arg Glu Ser Glu Val Ala Glu Leu 245 250 255 Thr Val Leu Glu Arg Pro Ser Phe Val Lys Arg Pro Ser Asn Leu Ala 260 265 270 Val Thr Val Asp Asp Ser Ala Glu Phe Lys Cys Glu Ala Arg Gly Asp 275 280 285 Pro Val Pro Thr Val Arg Trp Arg Lys Asp Asp Gly Glu Leu Pro Lys 290 295 300 Ser Arg Tyr Glu Ile Arg Asp Asp His Thr Leu Lys Ile Arg Lys Val 305 310 315 320 Thr Ala Gly Asp Met Gly Ser Tyr Thr Cys Val Ala Glu Asn Met Val 325 330 335 Gly Lys Ala Glu Ala Ser Ala Thr Leu Thr Val Gln Glu Pro Pro His 340 345 350 Phe Val Val Lys Pro Arg Asp Gln Val Val Ala Leu Gly Arg Thr Val 355 360 365 Thr Phe Gln Cys Glu Ala Thr Gly Asn Pro Gln Pro Ala Ile Phe Trp 370 375 380 Arg Arg Glu Gly Ser Gln Asn Leu Leu Phe Ser Tyr Gln Pro Pro Gln 385 390 395 400 Ser Ser Ser Phe Ser Val Ser Gln Thr Gly Asp Leu Thr Ile Thr 405 410 415 Asn Val Gln Arg Ser Asp Val Gly Tyr Tyr Ile Cys Gln Thr Leu Asn 420 425 430 Val Ala Gly Ser Ile Ile Thr Lys Ala Tyr Leu Glu Val Thr Asp Val 435 440 445 Ile Ala Asp Arg Pro Pro Pro Val Ile Arg Gln Gly Pro Val Asn Gln 450 455 460 Thr Val Ala Val Asp Gly Thr Phe Val Leu Ser Cys Val Ala Thr Gly 465 470 475 480 Ser Pro Val Pro Thr Ile Leu Trp Arg Lys Asp Gly Val Leu Val Ser 485 490 495 Thr Gln Asp Ser Arg Ile Lys Gln Leu Glu Asn Gly Val Leu Gln Ile 500 505 510 Arg Tyr Ala Lys Leu Gly Asp Thr Gly Arg Tyr Thr Cys Ile Ala Ser 515 520 525 Thr Pro Ser Gly Glu Ala Thr Trp Ser Ala Tyr Ile Glu Val Gln Glu 530 535 540 Phe Gly Val Pro Val Gln Pro Pro Arg Pro Thr Asp Pro Asn Leu Ile 545 550 555 560 Pro Ser Ala Pro Ser Lys Pro Glu Val Thr Asp Val Ser Arg Asn Thr 565 570 575 Val Thr Leu Ser Trp Gln Pro Asn Leu Asn Ser Gly Ala Thr Pro Thr 580 585 590 Ser Tyr Ile Ile Glu Ala Phe Ser His Ala Ser Gly Ser Ser Trp Gln 595 600 605 Thr Val Ala Glu Asn Val Lys Thr Glu Thr Ser Ala Ile Lys Gly Leu 610 615 620 Lys Pro Asn Ale Ile Tyr Leu Phe Leu Val Ala Ala Asn Ala Tyr 625 630 635 640 Gly Ile Ser Asp Pro Ser Gln Ile Ser Asp Pro Val Lys Thr Gln Asp 645 650 655 Val Leu Pro Thr Ser Gln Gly Val Asp His Lys Gln Val Gln Arg Glu 660 665 670 Leu Gly Asn Ala Val Leu His Leu His Asn Pro Thr Val Leu Ser Ser 675 680 685 Ser Ser Ile Glu Val His Trp Thr Val Asp Gln Gln Ser Gln Tyr Ile 690 695 700 Gln Gly Tyr Lys Ile Leu Tyr Arg Pro Ser Gly Ala Asn His Gly Glu 705 710 715 720 Ser Asp Trp Leu Val Phe Glu Val Arg Thr Pro Ala Lys Asn Ser Val 725 730 735 Val Ile Pro Asp Leu Arg Lys Gly Val Asn Tyr Glu Ile Lys Ala Arg 740 745 750 Pro Phe Phe Asn Glu Phe Gln Gly Ala Asp Ser Glu Ile Lys Phe Ala 755 760 765 Lys Thr Leu Glu Glu Ala Pro Ser Ala Pro Pro Gln Gly Val Thr Val 770 775 780 Ser Lys Asn Asp Gly Asn Gly Thr Ala Ile Leu Val Ser Trp Gln Pro 785 790 795 800 Pro Pro Glu Asp Thr Gln Asn Gly Met Val Gln Glu Tyr Lys Val Trp 805 810 815 Cys Leu Gly Asn Glu Thr Arg Tyr His Ile Asn Lys Thr Val Asp Gly 820 825 830 Ser Thr Phe Ser Val Valle Pro Phe Leu Val Pro Gly Ile Arg Tyr 835 840 845 Ser Val Glu Val Ala Ser Thr Gly Ala Gly Ser Gly Val Lys Ser 850 855 860 Glu Pro Gln Phe Ile Gln Leu Asp Ala His Gly Asn Pro Val Ser Pro 865 870 875 880 Glu Asp Gln Val Ser Leu Ala Gln Gln Ile Ser Asp Val Val Lys Gln 885 890 895 Pro Ala Phe Ile Ale Gly Ile Gly Ala Ala Cys Trp Ile Ile Leu Met 900 905 910 Val Phe Ser Ile Trp Leu Tyr Arg His Arg Lys Lys Arg Asn Gly Leu 915 920 925 Thr Ser Thr Tyr Ala Gly Ile Arg Lys Val Ser Ser Phe Thr Phe Thr 930 935 940 Pro Thr Val Thr Tyr Gln Arg Gly Gly Glu Ala Val Ser Ser Gly Gly 945 950 955 960 Arg Pro Gly Leu Leu Asn Ile Ser Glu Pro Ala Ala Gln Pro Trp Leu 965 970 975 Ala Asp Thr Trp Pro Asn Thr Gly Asn Asn His Asn Asp Cys Ser Ile 980 985 990 Ser Cys Cys Thr Ala Gly Asn Gly Asn Ser Asp Ser Asn Leu Thr Thr 995 1000 1005 Tyr Ser Arg Pro Ala Asp Cys Ile Ala Asn Tyr Asn Asn Gln Leu Asp 1010 1015 1020 Asn Lys Gln Thr Asn Leu Met Leu Pro Glu Ser Thr Val Tyr Gly Asp 1025 1030 1035 1040 Val Asp Leu Ser Asn Lys Ile Asn Glu Met Lys Thr Phe Asn Ser Pro 1045 1050 1055 Asn Leu Lys Asp Gly Arg Phe Val Asn Pro Ser Gly Gln Pro Thr Pro 1060 1065 1070 Tyr Ala Thr Thr Gln Leu Ile Gln Ser Asn Leu Ser Asn Asn Met Asn 1075 1080 1085 Asn Gly Ser Gly Asp Ser Gly Glu Lys His Trp Lys Pro Leu Gly Gln 1090 1095 1100 Gln Lys Gln Glu Val Ala Pro Val Gln Tyr Asn Ile Val Glu Gln Asn 1105 1110 1115 1120 Lys Leu Asn Lys Asp Tyr Arg Ala Asn Asp Thr Val Pro Thr Ile 1125 1130 1135 Pro Tyr Asn Gln Ser Tyr Asp Gln Asn Thr Gly Gly Ser Tyr Asn Ser 1140 1145 1150 Ser Asp Arg Gly Ser Ser Thr Ser Gly Ser Gln Gly His Lys Lys Gly 1155 1160 1165 Ala Arg Thr Pro Lys Val Pro Lys Gln Gly Gly Met Asn Trp Ala Asp 1170 1175 1180 Leu Leu Pro Pro Pro Pro Ala His Pro Pro Pro His Ser Asn Ser Glu 1185 1190 1195 1200 Glu Tyr Asn Ile Ser Val Asp Glu Ser Tyr Asp Gln Glu Met Pro Cys 1205 1210 1215 Pro Val Pro Pro Ala Arg Met Tyr Leu Gln Gln Asp Glu Leu Glu Glu 1220 1225 1230 Glu Glu Asp Glu Arg Gly Pro Thr Pro Pro Val Arg Gly Ala Ala Ser 1235 1240 1245 Ser Pro Ala Ala Val Ser Tyr Ser His Gln Ser Thr Ala Thr Leu Thr 1250 1255 1260 Pro Ser Pro Gln Glu Glu Leu Gln Pro Met Leu Gln Asp Cys Pro Glu 1265 1270 1275 1280 Glu Thr Gly His Met Gln His Gln Pro Asp Arg Arg Arg Gln Pro Val 1285 1290 1295 Ser Pro Pro Pro Pro Arg Pro Ile Ser Pro Pro His Thr Tyr Gly 1300 1305 1310 Tyr Ile Ser Gly Pro Leu Val Ser Asp Met Asp Thr Asp Ala Pro Glu 1315 1320 1325 Glu Glu Glu Asp Glu Ala Asp Met Glu Val Ala Lys Met Gln Thr Arg 1330 1335 1340 Arg Leu Leu Leu Arg Gly Leu Glu Gln Thr Pro Ala Ser Ser Val Gly 1345 1350 1355 1360 Asp Leu Glu Ser Ser Val Thr Gly Ser Met Ile Asn Gly Trp Gly Ser 1365 1370 1375 Ala Ser Glu Glu Asp Asn Ile Ser Ser Gly Arg Ser Ser Val Ser Ser 1380 1385 1390 Ser Asp Gly Ser Phe Phe Thr Asp Ala Asp Phe Ala Gln Ala Val Ala 1395 1400 1405 Ala Ala Ala Glu Tyr Ala Gly Leu Lys Val Ala Arg Arg Gln Met Gln 1410 1415 1420 Asp Ala Gly Arg Arg His Phe His Ala Ser Gln Cys Pro Arg Pro 1425 1430 1435 1440 Thr Ser Pro Val Ser Thr Asp Ser Asn Met Ser Ala Ala Val Met Gln 1445 1450 1455 Lys Thr Arg Pro Ala Lys Lys Leu Lys His Gln Pro Gly His Leu Arg 1460 1465 1470 Arg Glu Thr Tyr Thr Asp Asp Leu Pro Pro Pro Pro Val Pro Pro Pro 1475 1480 1485 Ala Ile Lys Ser Pro Thr Ala Gln Ser Lys Thr Gln Leu Glu Val Arg 1490 1495 1500 Pro Val Val Val Pro Lys Leu Pro Ser Met Asp Ala Arg Thr Asp Arg 1505 1510 1515 1520 Ser Ser Asp Arg Lys Gly Ser Ser Tyr Lys Gly Arg Glu Val Leu Asp 1525 1530 1535 Gly Arg Gln Val Val Asp Met Arg Thr Asn Pro Gly Asp Pro Arg Glu 1540 1545 1550 Ala Gln Glu Gln Gln Asn Asp Gly Lys Gly Arg Gly Asn Lys Ala Ala 1555 1560 1565 Lys Arg Asp Leu Pro Pro Ala Lys Thr His Leu Ile Gln Glu Asp Ile 1570 1575 1580 Leu Pro Tyr Cys Arg Pro Thr Phe Pro Thr Ser Asn Asn Pro Arg Asp 1585 1590 1595 1600 Pro Ser Ser Ser Ser Ser Met Ser Ser Arg Gly Ser Gly Ser Arg Gln 1605 1610 1615 Arg Glu Gln Ala Asn Val Gly Arg Arg Asn Ile Ala Glu Met Gln Val 1620 1625 1630 Leu Gly Gly Tyr Glu Arg Gly Glu Asp Asn Asn Glu Glu Leu Glu Glu 1635 1640 1645 Thr Glu Ser 1650 <210> 8 <211> 1606 <212> PRT <213> roundabout homolog 1 isoform b [Homo sapiens] <400> 8 Met Ile Ala Glu Pro Ala His Phe Tyr Leu Phe Gly Leu Ile Cys Leu 1 5 10 15 Cys Ser Gly Ser Arg Leu Arg Gln Glu Asp Phe Pro Pro Arg Ile Val 20 25 30 Glu His Pro Ser Asp Leu Ile Val Ser Lys Gly Glu Pro Ala Thr Leu 35 40 45 Asn Cys Lys Ala Glu Gly Arg Pro Thr Pro Thr Ile Glu Trp Tyr Lys 50 55 60 Gly Gly Glu Arg Val Glu Thr Asp Lys Asp Asp Pro Arg Ser His Arg 65 70 75 80 Met Leu Leu Pro Ser Gly Ser Leu Phe Phe Leu Arg Ile Val His Gly 85 90 95 Arg Lys Ser Arg Pro Asp Glu Gly Val Tyr Val Cys Val Ala Arg Asn 100 105 110 Tyr Leu Gly Glu Glu Ala Val Ser As Asn Ala Ser Leu Glu Val Ala Ile 115 120 125 Leu Arg Asp Asp Phe Arg Gln Asn Pro Ser Asp Val Met Val Ala Val 130 135 140 Gly Glu Pro Ala Val Met Glu Cys Gln Pro Pro Arg Gly His Pro Glu 145 150 155 160 Pro Thr Ile Ser Trp Lys Lys Asp Gly Ser Pro Leu Asp Asp Lys Asp 165 170 175 Glu Arg Ile Thr Ile Arg Gly Gly Lys Leu Met Ile Thr Tyr Thr Arg 180 185 190 Lys Ser Asp Ala Gly Lys Tyr Val Cys Val Gly Thr Asn Met Val Gly 195 200 205 Glu Arg Glu Ser Glu Val Ala Glu Leu Thr Val Leu Glu Arg Pro Ser 210 215 220 Phe Val Lys Arg Pro Ser Asn Leu Ala Val Thr Val Asp Asp Ser Ala 225 230 235 240 Glu Phe Lys Cys Glu Ala Arg Gly Asp Pro Val Val Thr Val Arg Trp 245 250 255 Arg Lys Asp Asp Gly Glu Leu Pro Lys Ser Arg Tyr Glu Ile Arg Asp 260 265 270 Asp His Thr Leu Lys Ile Arg Lys Val Thr Ala Gly Asp Met Gly Ser 275 280 285 Tyr Thr Cys Val Ala Glu Asn Met Val Gly Lys Ala Glu Ala Ser Ala 290 295 300 Thr Leu Thr Val Gln Val Gly Ser Glu Pro Pro His Phe Val Val Lys 305 310 315 320 Pro Arg Asp Gln Val Val Ala Leu Gly Arg Thr Val Thr Phe Gln Cys 325 330 335 Glu Ala Thr Gly Asn Pro Gln Pro Ala Ile Phe Trp Arg Arg Glu Gly 340 345 350 Ser Gln Asn Leu Leu Phe Ser Tyr Gln Pro Pro Gln Ser Ser Ser Arg 355 360 365 Phe Ser Val Ser Gln Thr Gly Asp Leu Thr Ile Thr Asn Val Gln Arg 370 375 380 Ser Asp Val Gly Tyr Tyr Ile Cys Gln Thr Leu Asn Val Ala Gly Ser 385 390 395 400 Ile Ile Thr Lys Ala Tyr Leu Glu Val Thr Asp Val Ile Ala Asp Arg 405 410 415 Pro Pro Pro Val Ile Arg Gln Gly Pro Val Asn Gln Thr Val Ala Val 420 425 430 Asp Gly Thr Phe Val Leu Ser Cys Val Ala Thr Gly Ser Pro Val Pro 435 440 445 Thr Ile Leu Trp Arg Lys Asp Gly Val Leu Val Ser Thr Gln Asp Ser 450 455 460 Arg Ile Lys Gln Leu Glu Asn Gly Val Leu Gln Ile Arg Tyr Ala Lys 465 470 475 480 Leu Gly Asp Thr Gly Arg Tyr Thr Cys Ile Ala Ser Thr Pro Ser Gly 485 490 495 Glu Ala Thr Trp Ser Ala Tyr Ile Glu Val Gln Glu Phe Gly Val Pro 500 505 510 Val Gln Pro Pro Arg Pro Thr Asp Pro Asn Leu Ile Pro Ser Ala Pro 515 520 525 Ser Lys Pro Glu Val Thr Asp Val Ser Arg Asn Thr Val Thr Leu Ser 530 535 540 Trp Gln Pro Asn Leu Asn Ser Gly Ala Thr Pro Thr Ser Tyr Ile Ile 545 550 555 560 Glu Ala Phe Ser Ala Ser Gly Ser Ser Trp Gln Thr Val Ala Glu 565 570 575 Asn Val Lys Thr Glu Thr Ser Ala Ile Lys Gly Leu Lys Pro Asn Ala 580 585 590 Ile Tyr Leu Phe Leu Val Arg Ala Asn Ala Tyr Gly Ile Ser Asp 595 600 605 Pro Ser Gln Ile Ser Asp Pro Val Lys Thr Gln Asp Val Leu Pro Thr 610 615 620 Ser Gln Gly Val Asp His Lys Gln Val Gln Arg Glu Leu Gly Asn Ala 625 630 635 640 Val Leu His Leu His Asn Pro Thr Val Leu Ser Ser Ser Ser Ile Glu 645 650 655 Val His Trp Thr Val Asp Gln Gln Ser Gln Tyr Ile Gln Gly Tyr Lys 660 665 670 Ile Leu Tyr Arg Pro Ser Gly Ala Asn His Gly Glu Ser Asp Trp Leu 675 680 685 Val Phe Glu Val Arg Thr Pro Ala Lys Asn Ser Val Val Ile Pro Asp 690 695 700 Leu Arg Lys Gly Val Asn Tyr Glu Ile Lys Ala Arg Pro Phe Phe Asn 705 710 715 720 Glu Phe Gln Gly Ala Asp Ser Glu Ile Lys Phe Ala Lys Thr Leu Glu 725 730 735 Glu Ala Pro Ser Ala Pro Pro Gln Gly Val Thr Val Ser Lys Asn Asp 740 745 750 Gly Asn Gly Thr Ala Ile Leu Val Ser Trp Gln Pro Pro Pro Glu Asp 755 760 765 Thr Gln Asn Gly Met Val Gln Glu Tyr Lys Val Trp Cys Leu Gly Asn 770 775 780 Glu Thr Arg Tyr His Ile Asn Lys Thr Val Asp Gly Ser Thr Phe Ser 785 790 795 800 Val Valle Pro Phe Leu Val Pro Gly Ile Arg Tyr Ser Val Glu Val 805 810 815 Ala Ala Ser Thr Gly Ala Gly Ser Gly Val Lys Ser Glu Pro Gln Phe 820 825 830 Ile Gln Leu Asp Ala His Gly Asn Pro Val Ser Pro Glu Asp Gln Val 835 840 845 Ser Leu Ala Gln Gln Ile Ser Asp Val Val Lys Gln Pro Ala Phe Ile 850 855 860 Ala Gly Ile Gly Ala Ala Cys Trp Ile Ile Leu Met Val Phe Ser Ile 865 870 875 880 Trp Leu Tyr Arg His Arg Lys Lys Arg Asn Gly Leu Thr Ser Thr Tyr 885 890 895 Ala Gly Ile Arg Lys Val Thr Tyr Gln Arg Gly Gly Glu Ala Val Ser 900 905 910 Ser Gly Gly Arg Pro Gly Leu Leu Asn Ile Ser Glu Pro Ala Ala Gln 915 920 925 Pro Trp Leu Ala Asp Thr Trp Pro Asn Thr Gly Asn Asn His Asn Asp 930 935 940 Cys Ser Ile Ser Cys Cys Thr Ala Gly Asn Gly Asn Ser Asp Ser Asn 945 950 955 960 Leu Thr Thr Ser Ser Arg Pro Ala Asp Cys Ile Ala Asn Tyr Asn Asn 965 970 975 Gln Leu Asp Asn Lys Gln Thr Asn Leu Met Leu Pro Glu Ser Thr Val 980 985 990 Tyr Gly Asp Val Asp Leu Ser Asn Lys Ile Asn Glu Met Lys Thr Phe 995 1000 1005 Asn Ser Pro Asn Leu Lys Asp Gly Arg Phe Val Asn Pro Ser Gly Gln 1010 1015 1020 Pro Thr Pro Tyr Ala Thr Thr Gln Leu Ile Gln Ser Asn Leu Ser Asn 1025 1030 1035 1040 Asn Met Asn Asn Gly Ser Gly Asp Ser Gly Glu Lys His Trp Lys Pro 1045 1050 1055 Leu Gly Gln Gln Lys Gln Glu Val Ala Pro Val Gln Tyr Asn Ile Val 1060 1065 1070 Glu Gln Asn Lys Leu Asn Lys Asp Tyr Arg Ala Asn Asp Thr Val Pro 1075 1080 1085 Pro Thr Ile Pro Tyr Asn Gln Ser Tyr Asp Gln Asn Thr Gly Gly Ser 1090 1095 1100 Tyr Asn Ser Ser Asp Arg Gly Ser Ser Thr Ser Gly Ser Gln Gly His 1105 1110 1115 1120 Lys Lys Gly Ala Arg Thr Pro Lys Val Pro Lys Gln Gly Gly Met Asn 1125 1130 1135 Trp Ala Asp Leu Leu Pro Pro Pro Pro Ala His Pro Pro Pro His Ser 1140 1145 1150 Asn Ser Glu Glu Tyr Asn Ile Ser Val Asp Glu Ser Tyr Asp Gln Glu 1155 1160 1165 Met Pro Cys Pro Val Pro Pro Ala Arg Met Tyr Leu Gln Gln Asp Glu 1170 1175 1180 Leu Glu Glu Glu Glu Asp Glu Arg Gly Pro Thr Pro Pro Val Val Gly 1185 1190 1195 1200 Ala Ala Ser Ser Ala Ala Val Ser Tyr Ser His Gln Ser Thr Ala 1205 1210 1215 Thr Leu Thr Pro Ser Pro Gln Glu Glu Leu Gln Pro Met Leu Gln Asp 1220 1225 1230 Cys Pro Glu Glu Thr Gly His Met Gln His Gln Pro Asp Arg Arg Arg 1235 1240 1245 Gln Pro Val Ser Pro Pro Pro Pro Pro Arg Pro Ile Ser Pro Pro His 1250 1255 1260 Thr Tyr Gly Tyr Ile Ser Gly Pro Leu Val Ser Asp Met Asp Thr Asp 1265 1270 1275 1280 Ala Pro Glu Glu Glu Glu Glu Asp Glu Ala Asp Met Glu Val Ala Lys Met 1285 1290 1295 Gln Thr Arg Arg Leu Leu Leu Arg Gly Leu Glu Gln Thr Pro Ala Ser 1300 1305 1310 Ser Val Gly Asp Leu Glu Ser Ser Val Thr Gly Ser Ser Ile Asn Gly 1315 1320 1325 Trp Gly Ser Ala Ser Glu Glu Asp Asn Ile Ser Ser Gly Arg Ser Ser 1330 1335 1340 Val Ser Ser Ser Asp Gly Ser Phe Phe Thr Asp Ala Asp Phe Ala Gln 1345 1350 1355 1360 Ala Val Ala Ala Ala Glu Tyr Ala Gly Leu Lys Val Ala Arg Arg 1365 1370 1375 Gln Met Gln Asp Ala Ala Gly Arg Arg His Phe His Ala Ser Gln Cys 1380 1385 1390 Pro Arg Pro Thr Ser Pro Val Ser Thr Asp Ser Asn Met Ser Ala Ala 1395 1400 1405 Val Met Gln Lys Thr Arg Pro Ala Lys Lys Leu Lys His Gln Pro Gly 1410 1415 1420 His Leu Arg Arg Glu Thr Tyr Thr Asp Asp Leu Pro Pro Pro Pro Val 1425 1430 1435 1440 Pro Pro Pro Ala Ile Lys Ser Pro Thr Ala Gln Ser Lys Thr Gln Leu 1445 1450 1455 Glu Val Arg Pro Val Val Val Pro Lys Leu Pro Ser Met Asp Ala Arg 1460 1465 1470 Thr Asp Arg Ser Ser Asp Arg Lys Gly Ser Ser Tyr Lys Gly Arg Glu 1475 1480 1485 Val Leu Asp Gly Arg Gln Val Val Asp Met Arg Thr Asn Pro Gly Asp 1490 1495 1500 Pro Arg Glu Ala Gln Glu Gln Gln Asn Asp Gly Lys Gly Arg Gly Asn 1505 1510 1515 1520 Lys Ala Ala Lys Arg Asp Leu Pro Pro Ala Lys Thr His Leu Ile Gln 1525 1530 1535 Glu Asp Ile Leu Pro Tyr Cys Arg Pro Thr Phe Pro Thr Ser Asn Asn 1540 1545 1550 Pro Arg Asp Pro Ser Ser Ser Ser Ser Met Ser Ser Arg Gly Ser Gly 1555 1560 1565 Ser Arg Gln Arg Glu Gln Ala Asn Val Gly Arg Arg Asn Ile Ala Glu 1570 1575 1580 Met Gln Val Leu Gly Gly Tyr Glu Arg Gly Glu Asp Asn Asn Glu Glu 1585 1590 1595 1600 Leu Glu Glu Thr Glu Ser 1605 <210> 9 <211> 1551 <212> PRT <213> roundabout homolog 1 isoform d [Homo sapiens] <400> 9 Met Ile Ala Glu Pro Ala His Phe Tyr Leu Phe Gly Leu Ile Cys Leu 1 5 10 15 Cys Ser Gly Ser Arg Leu Arg Gln Glu Asp Phe Pro Pro Arg Ile Val 20 25 30 Glu His Pro Ser Asp Leu Ile Val Ser Lys Gly Glu Pro Ala Thr Leu 35 40 45 Asn Cys Lys Ala Glu Gly Arg Pro Thr Pro Thr Ile Glu Trp Tyr Lys 50 55 60 Gly Gly Glu Arg Val Glu Thr Asp Lys Asp Asp Pro Arg Ser His Arg 65 70 75 80 Met Leu Leu Pro Ser Gly Ser Leu Phe Phe Leu Arg Ile Val His Gly 85 90 95 Arg Lys Ser Arg Pro Asp Glu Gly Val Tyr Val Cys Val Ala Arg Asn 100 105 110 Tyr Leu Gly Glu Glu Ala Val Ser As Asn Ala Ser Leu Glu Val Ala Ile 115 120 125 Leu Arg Asp Asp Phe Arg Gln Asn Pro Ser Asp Val Met Val Ala Val 130 135 140 Gly Glu Pro Ala Val Met Glu Cys Gln Pro Pro Arg Gly His Pro Glu 145 150 155 160 Pro Thr Ile Ser Trp Lys Lys Asp Gly Ser Pro Leu Asp Asp Lys Asp 165 170 175 Glu Arg Ile Thr Ile Arg Gly Gly Lys Leu Met Ile Thr Tyr Thr Arg 180 185 190 Lys Ser Asp Ala Gly Lys Tyr Val Cys Val Gly Thr Asn Met Val Gly 195 200 205 Glu Arg Glu Ser Glu Val Ala Glu Leu Thr Val Leu Glu Arg Pro Ser 210 215 220 Phe Val Lys Arg Pro Ser Asn Leu Ala Val Thr Val Asp Asp Ser Ala 225 230 235 240 Glu Phe Lys Cys Glu Ala Arg Gly Asp Pro Val Val Thr Val Arg Trp 245 250 255 Arg Lys Asp Asp Gly Glu Leu Pro Lys Ser Arg Tyr Glu Ile Arg Asp 260 265 270 Asp His Thr Leu Lys Ile Arg Lys Val Thr Ala Gly Asp Met Gly Ser 275 280 285 Tyr Thr Cys Val Ala Glu Asn Met Val Gly Lys Ala Glu Ala Ser Ala 290 295 300 Thr Leu Thr Val Gln Val Gly Ser Glu Pro Pro His Phe Val Val Lys 305 310 315 320 Pro Arg Asp Gln Val Val Ala Leu Gly Arg Thr Val Thr Phe Gln Cys 325 330 335 Glu Ala Thr Gly Asn Pro Gln Pro Ala Ile Phe Trp Arg Arg Glu Gly 340 345 350 Ser Gln Asn Leu Leu Phe Ser Tyr Gln Pro Pro Gln Ser Ser Ser Arg 355 360 365 Phe Ser Val Ser Gln Thr Gly Asp Leu Thr Ile Thr Asn Val Gln Arg 370 375 380 Ser Asp Val Gly Tyr Tyr Ile Cys Gln Thr Leu Asn Val Ala Gly Ser 385 390 395 400 Ile Ile Thr Lys Ala Tyr Leu Glu Val Thr Asp Val Ile Ala Asp Arg 405 410 415 Pro Pro Pro Val Ile Arg Gln Gly Pro Val Asn Gln Thr Val Ala Val 420 425 430 Asp Gly Thr Phe Val Leu Ser Cys Val Ala Thr Gly Ser Pro Val Pro 435 440 445 Thr Ile Leu Trp Arg Lys Asp Gly Val Leu Val Ser Thr Gln Asp Ser 450 455 460 Arg Ile Lys Gln Leu Glu Asn Gly Val Leu Gln Ile Arg Tyr Ala Lys 465 470 475 480 Leu Gly Asp Thr Gly Arg Tyr Thr Cys Ile Ala Ser Thr Pro Ser Gly 485 490 495 Glu Ala Thr Trp Ser Ala Tyr Ile Glu Val Gln Glu Phe Gly Val Pro 500 505 510 Val Gln Pro Pro Arg Pro Thr Asp Pro Asn Leu Ile Pro Ser Ala Pro 515 520 525 Ser Lys Pro Glu Val Thr Asp Val Ser Arg Asn Thr Val Thr Leu Ser 530 535 540 Trp Gln Pro Asn Leu Asn Ser Gly Ala Thr Pro Thr Ser Tyr Ile Ile 545 550 555 560 Glu Ala Phe Ser Ala Ser Gly Ser Ser Trp Gln Thr Val Ala Glu 565 570 575 Asn Val Lys Thr Glu Thr Ser Ala Ile Lys Gly Leu Lys Pro Asn Ala 580 585 590 Ile Tyr Leu Phe Leu Val Arg Ala Asn Ala Tyr Gly Ile Ser Asp 595 600 605 Pro Ser Gln Ile Ser Asp Pro Val Lys Thr Gln Asp Val Leu Pro Thr 610 615 620 Ser Gln Gly Val Asp His Lys Gln Val Gln Arg Glu Leu Gly Asn Ala 625 630 635 640 Val Leu His Leu His Asn Pro Thr Val Leu Ser Ser Ser Ser Ile Glu 645 650 655 Val His Trp Thr Val Asp Gln Gln Ser Gln Tyr Ile Gln Gly Tyr Lys 660 665 670 Ile Leu Tyr Arg Pro Ser Gly Ala Asn His Gly Glu Ser Asp Trp Leu 675 680 685 Val Phe Glu Val Arg Thr Pro Ala Lys Asn Ser Val Val Ile Pro Asp 690 695 700 Leu Arg Lys Gly Val Asn Tyr Glu Ile Lys Ala Arg Pro Phe Phe Asn 705 710 715 720 Glu Phe Gln Gly Ala Asp Ser Glu Ile Lys Phe Ala Lys Thr Leu Glu 725 730 735 Glu Ala Pro Ser Ala Pro Pro Gln Gly Val Thr Val Ser Lys Asn Asp 740 745 750 Gly Asn Gly Thr Ala Ile Leu Val Ser Trp Gln Pro Pro Pro Glu Asp 755 760 765 Thr Gln Asn Gly Met Val Gln Glu Tyr Lys Val Trp Cys Leu Gly Asn 770 775 780 Glu Thr Arg Tyr His Ile Asn Lys Thr Val Asp Gly Ser Thr Phe Ser 785 790 795 800 Val Valle Pro Phe Leu Val Pro Gly Ile Arg Tyr Ser Val Glu Val 805 810 815 Ala Ala Ser Thr Gly Ala Gly Ser Gly Val Lys Ser Glu Pro Gln Phe 820 825 830 Ile Gln Leu Asp Ala His Gly Asn Pro Val Ser Pro Glu Asp Gln Val 835 840 845 Ser Leu Ala Gln Gln Ile Ser Asp Val Val Lys Gln Pro Ala Phe Ile 850 855 860 Ala Gly Ile Gly Ala Ala Cys Trp Ile Ile Leu Met Val Phe Ser Ile 865 870 875 880 Trp Leu Tyr Arg His Arg Lys Lys Arg Asn Gly Leu Thr Ser Thr Tyr 885 890 895 Ala Gly Ile Arg Lys Val Thr Tyr Gln Arg Gly Gly Glu Ala Val Ser 900 905 910 Ser Gly Gly Arg Pro Gly Leu Leu Asn Ile Ser Glu Pro Ala Ala Gln 915 920 925 Pro Trp Leu Ala Asp Thr Trp Pro Asn Thr Gly Asn Asn His Asn Asp 930 935 940 Cys Ser Ile Ser Cys Cys Thr Ala Gly Asn Gly Asn Ser Asp Ser Asn 945 950 955 960 Leu Thr Thr Tyr Ser Arg Pro Gly Gln Pro Thr Pro Tyr Ala Thr Thr 965 970 975 Gln Leu Ile Gln Ser Asn Leu Ser Asn Asn Met Asn Asn Gly Ser Gly 980 985 990 Asp Ser Gly Glu Lys His Trp Lys Pro Leu Gly Gln Gln Lys Gln Glu 995 1000 1005 Val Ala Pro Val Gln Tyr Asn Ile Val Glu Gln Asn Lys Leu Asn Lys 1010 1015 1020 Asp Tyr Arg Ala Asn Asp Thr Val Pro Pro Thr Ile Pro Tyr Asn Gln 1025 1030 1035 1040 Ser Tyr Asp Gln Asn Thr Gly Gly Ser Tyr Asn Ser Ser Asp Arg Gly 1045 1050 1055 Ser Ser Thr Ser Gly Ser Gln Gly His Lys Lys Gly Ala Arg Thr Pro 1060 1065 1070 Lys Val Pro Lys Gln Gly Gly Met Asn Trp Ala Asp Leu Leu Pro Pro 1075 1080 1085 Pro Pro Ala His Pro Pro Pro His Ser Asn Ser Glu Glu Tyr Asn Ile 1090 1095 1100 Ser Val Asp Glu Ser Tyr Asp Gln Glu Met Pro Cys Pro Val Pro Pro 1105 1110 1115 1120 Ala Arg Met Tyr Leu Gln Gln Asp Glu Leu Glu Glu Glu Glu Asp Glu 1125 1130 1135 Arg Gly Pro Thr Pro Pro Val Val Gly Ala Ala Ser Ser Ala Ala 1140 1145 1150 Val Ser Tyr Ser His Gln Ser Thr Ala Thr Leu Thr Ser Ser Pro Gln 1155 1160 1165 Glu Glu Leu Gln Pro Met Leu Gln Asp Cys Pro Glu Glu Thr Gly His 1170 1175 1180 Met Gln His Gln Pro Asp Arg Arg Arg Gln Pro Val Ser Pro Pro Pro 1185 1190 1195 1200 Pro Pro Arg Pro Ile Ser Pro Pro His Thr Tyr Gly Tyr Ile Ser Gly 1205 1210 1215 Pro Leu Val Ser Asp Met Asp Thr Asp Ala Pro Glu Glu Glu Glu Asp 1220 1225 1230 Glu Ala Asp Met Glu Val Ala Lys Met Gln Thr Arg Arg Leu Leu Leu 1235 1240 1245 Arg Gly Leu Glu Gln Thr Pro Ala Ser Ser Val Gly Asp Leu Glu Ser 1250 1255 1260 Ser Val Thr Gly Ser Met Ile Asn Gly Trp Gly Ser Ala Ser Glu Glu 1265 1270 1275 1280 Asp Asn Ile Ser Ser Gly Arg Ser Ser Val Ser Ser Ser Asp Gly Ser 1285 1290 1295 Phe Phe Thr Asp Ala Asp Phe Ala Gln Ala Val Ala Ala Ala Ala Glu 1300 1305 1310 Tyr Ala Gly Leu Lys Val Ala Arg Arg Gln Met Gln Asp Ala Ala Gly 1315 1320 1325 Arg Arg His Phe His Ala Ser Gln Cys Pro Arg Pro Thr Ser Pro Val 1330 1335 1340 Ser Thr Asp Ser Asn Met Ser Ala Ala Val Met Gln Lys Thr Arg Pro 1345 1350 1355 1360 Ala Lys Lys Leu Lys His Gln Pro Gly His Leu Arg Arg Glu Thr Tyr 1365 1370 1375 Thr Asp Leu Pro Pro Pro Pro Val Pro Pro Pro Ala Ile Lys Ser 1380 1385 1390 Pro Thr Ala Gln Ser Lys Thr Gln Leu Glu Val Val Pro Val Val Val 1395 1400 1405 Pro Lys Leu Pro Ser Met Asp Ala Arg Thr Asp Arg Ser Ser Asp Arg 1410 1415 1420 Lys Gly Ser Ser Tyr Lys Gly Arg Glu Val Leu Asp Gly Arg Gln Val 1425 1430 1435 1440 Val Asp Met Arg Thr Asn Pro Gly Asp Pro Arg Glu Ala Gln Glu Gln 1445 1450 1455 Gln Asn Asp Gly Lys Gly Arg Gly Asn Lys Ala Ala Lys Arg Asp Leu 1460 1465 1470 Pro Pro Ala Lys Thr His Leu Ile Gln Glu Asp Ile Leu Pro Tyr Cys 1475 1480 1485 Arg Pro Thr Phe Pro Thr Ser Asn Asn Pro Arg Asp Pro Ser Ser Ser 1490 1495 1500 Ser Ser Met Ser Ser Arg Gly Ser Gly Ser Arg Gln Arg Glu Gln Ala 1505 1510 1515 1520 Asn Val Gly Arg Arg Asn Ile Ala Glu Met Gln Val Leu Gly Gly Tyr 1525 1530 1535 Glu Arg Gly Glu Asp Asn Asn Glu Glu Leu Glu Glu Thr Glu Ser 1540 1545 1550 <210> 10 <211> 6895 <212> DNA <213> Homo sapiens roundabout, axon guidance receptor, homolog 1 (Drosophila) (ROBO1) <400> 10 cccgacttca ctctctccct atttccccac tcttaggttt aaaagtctgt cacctttcgc 60 ttggtttaaa ctcggaaagg tctcagtgca cagcaaagtt gcagggctgc gtctgcacta 120 cggagcctct agattgctga aaacagtctt atggaaggat aacacattgt ctgtcactgg 180 ctggttgtaa tgcaaggaag ggacaaagat gaaatggaaa catgttcctt ttttggtcat 240 gatatcactc ctcagcttat ccccaaatca cctgtttctg gcccagctta ttccagaccc 300 tgaagatgta gagaggggga acgaccacgg gacgccaatc cccacctctg ataacgatga 360 caattcgctg ggctatacag gctcccgtct tcgtcaggaa gattttccac ctcgcattgt 420 tgaacaccct tcagacctga ttgtctcaaa aggagaacct gcaactttga actgcaaagc 480 tgaaggccgc cccacaccca ctattgaatg gtacaaaggg ggagagagag tggagacaga 540 caaagatgac cctcgctcac accgaatgtt gctgccgagt ggatctttat ttttcttacg 600 tatagtacat ggacggaaaa gtagacctga tgaaggagtc tatgtctgtg tagcaaggaa 660 ttaccttgga gaggctgtga gccacaatgc atcgctggaa gtagccatac ttcgggatga 720 cttcagacaa aacccttcgg atgtcatggt tgcagtagga gagcctgcag taatggaatg 780 ccaacctcca cgaggccatc ctgagcccac catttcatgg aagaaagatg gctctccact 840 ggatgataaa gatgaaagaa taactatacg aggaggaaag ctcatgatca cttacacccg 900 taaaagtgac gctggcaaat atgtttgtgt tggtaccaat atggttgggg aacgtgagag 960 tgaagtagcc gagctgactg tcttagagag accatcattt gtgaagagac ccagtaactt 1020 ggcagtaact gtggatgaca gtgcagaatt taaatgtgag gcccgaggtg accctgtacc 1080 tacagtacga tggaggaaag atgatggaga gctgcccaaa tccagatatg aaatccgaga 1140 tgatcatacc ttgaaaatta ggaaggtgac agctggtgac atgggttcat acacttgtgt 1200 tgcagaaaat atggtgggca aagctgaagc atctgctact ctgactgttc aagaacctcc 1260 acattttgtt gtgaaacccc gtgaccaggt tgttgctttg ggacggactg taacttttca 1320 gtgtgaagca accggaaatc ctcaaccagc tattttctgg aggagagaag ggagtcagaa 1380 tctacttttc tcatatcaac caccacagtc atccagccga ttttcagtct cccagactgg 1440 cgacctcaca attactaatg tccagcgatc tgatgttggt tattacatct gccagacttt 1500 aaatgttgct ggaagcatca tcacaaaggc atatttggaa gttacagatg tgattgcaga 1560 tcggcctccc ccagttattc gacaaggtcc tgtgaatcag actgtagccg tggatggcac 1620 tttcgtcctc agctgtgtgg ccacaggcag tccagtgccc accattctgt ggagaaagga 1680 tggagtcctc gtttcaaccc aagactctcg aatcaaacag ttggagaatg gagtactgca 1740 gatccgatat gctaagctgg gtgatactgg tcggtacacc tgcattgcat caacccccag 1800 tggtgaagca acatggagtg cttacattga agttcaagaa tttggagttc cagttcagcc 1860 tccaagacct actgacccaa atttaatccc tagtgcccca tcaaaacctg aagtgacaga 1920 tgtcagcaga aatacagtca cattatcgtg gcaaccaaat ttgaattcag gagcaactcc 1980 aacatcttat attatagaag ccttcagcca tgcatctggt agcagctggc agaccgtagc 2040 agagaatgtg aaaacagaaa catctgccat taaaggactc aaacctaatg caatttacct 2100 tttccttgtg agggcagcta atgcatatgg aattagtgat ccaagccaaa tatcagatcc 2160 agtgaaaaca caagatgtcc taccaacaag tcagggggtg gaccacaagc aggtccagag 2220 agagctggga aatgctgttc tgcacctcca caaccccacc gtcctttctt cctcttccat 2280 cgaagtgcac tggacagtag atcaacagtc tcagtatata caaggatata aaattctcta 2340 tcggccatct ggagccaacc acggagaatc agactggtta gtttttgaag tgaggacgcc 2400 agccaaaaac agtgtggtaa tccctgatct cagaaaggga gtcaactatg aaattaaggc 2460 tcgccctttt tttaatgaat ttcaaggagc agatagtgaa atcaagtttg ccaaaaccct 2520 ggaagaagca cccagtgccc caccccaagg tgtaactgta tccaagaatg atggaaacgg 2580 aactgcaatt ctagttagtt ggcagccacc tccagaagac actcaaaatg gaatggtcca 2640 agagtataag gtttggtgtc tgggcaatga aactcgatac cacatcaaca aaacagtgga 2700 tggttccacc ttttccgtgg tcattccctt tcttgttcct ggaatccgat acagtgtgga 2760 agtggcagcc agcactgggg ctgggtctgg ggtaaagagt gagcctcagt tcatccagct 2820 ggatgcccat ggaaaccctg tgtcacctga ggaccaagtc agcctcgctc agcagatttc 2880 agatgtggtg aagcagccgg ccttcatagc aggtattgga gcagcctgtt ggatcatcct 2940 catggtcttc agcatctggc tttatcgaca ccgcaagaag agaaacggac ttactagtac 3000 ctacgcgggt atcagaaaag tcccgtcttt taccttcaca ccaacagtaa cttaccagag 3060 aggaggcgaa gctgtcagca gtggagggag gcctggactt ctcaacatca gtgaacctgc 3120 cgcgcagcca tggctggcag acacgtggcc taatactggc aacaaccaca atgactgctc 3180 catcagctgc tgcacggcag gcaatggaaa cagcgacagc aacctcacta cctacagtcg 3240 cccagctgat tgtatagcaa attataacaa ccaactggat aacaaacaaa caaatctgat 3300 gctccctgag tcaactgttt atggtgatgt ggaccttagt aacaaaatca atgagatgaa 3360 aaccttcaat agcccaaatc tgaaggatgg gcgttttgtc aatccatcag ggcagcctac 3420 tccttacgcc accactcagc tcatccagtc aaacctcagc aacaacatga acaatggcag 3480 cggggactct ggcgagaagc actggaaacc actgggacag cagaaacaag aagtggcacc 3540 agttcagtac aacatcgtgg agcaaaacaa gctgaacaaa gattatcgag caaatgacac 3600 agttcctcca actatcccat acaaccaatc atacgaccag aacacaggag gatcctacaa 3660 cagctcagac cggggcagta gtacatctgg gagtcagggg cacaagaaag gggcaagaac 3720 ctccccagc 3780 acatcctcct ccacacagca atagcgaaga gtacaacatt tctgtagatg aaagctatga 3840 ccaagaaatg ccatgtcccg tgccaccagc aaggatgtat ttgcaacaag atgaattaga 3900 agaggaggaa gatgaacgag gccccactcc ccctgttcgg ggagcagctt cttctccagc 3960 tgccgtgtcc tatagccatc agtccactgc cactctgact ccctccccac aggaagaact 4020 ccagcccatg ttacaggatt gtccagagga gactggccac atgcagcacc agcccgacag 4080 gagacggcag cctgtgagtc ctcctccacc accacggccg atctcccctc cacataccta 4140 tggctacatt tcaggacccc tggtctcaga tatggatacg gatgcgccag aagaggaaga 4200 agacgaagcc gacatggagg tagccaagat gcaaaccaga aggcttttgt tacgtgggct 4260 tgagcagaca cctgcctcca gtgttgggga cctggagagc tctgtcacgg ggtccatgat 4320 caacggctgg ggctcagcct cagaggagga caacatttcc agcggacgct ccagtgttag 4380 ttcttcggac ggctcctttt tcactgatgc tgactttgcc caggcagtcg cagcagcggc 4440 agagtatgct ggtctgaaag tagcacgacg gcaaatgcag gatgctgctg gccgtcgaca 4500 ttttcatgcg tctcagtgcc ctaggcccac aagtcccgtg tctacagaca gcaacatgag 4560 tgccgccgta atgcagaaaa ccagaccagc caagaaactg aaacaccagc caggacatct 4620 gcgcagagaa acctacacag atgatcttcc accacctcct gtgccgccac ctgctataaa 4680 gtcacctact gcccaatcca agacacagct ggaagtacga cctgtagtgg tgccaaaact 4740 cccttctatg gatgcaagaa cagacagatc atcagacaga aaaggaagca gttacaaggg 4800 gagagaagtg ttggatggaa gacaggttgt tgacatgcga acaaatccag gtgatcccag 4860 agaagcacag gaacagcaaa atgacgggaa aggacgtgga aacaaggcag caaaacgaga 4920 ccttccacca gcaaagactc atctcatcca agaggatatt ctaccttatt gtagacctac 4980 ttttccaaca tcaaataatc ccagagatcc cagttcctca agctcaatgt catcaagagg 5040 atcaggaagc agacaaagag aacaagcaaa tgtaggtcga agaaatattg cagaaatgca 5100 ggtacttgga ggatatgaaa gaggagaaga taataatgaa gaattagagg aaactgaaag 5160 ctgaagacaa ccaagaggct tatgagatct aatgtgaaaa tcatcactca agatgcctcc 5220 tgtcagatga cacatgacgc cagataaaat gttcagtgca atcagagtgt acaaattgtc 5280 gtttttattc ctcttattgg gatatcattt taaaaacttt attgggtttt tattgttgtt 5340 gtttgatccc taaccctaca aagagccttc ctattcccct cgctgttgga gcaaaccatt 5400 ataccttact tccagcaagc aaagtgcttt gacttcttgc ttcagtcatc agccagcaag 5460 agggaacaaa actgttcttt tgcattttgc cgctgagata tggcattgca ctgcttatat 5520 gccaagctaa tttatagcaa gatattgatc aaatatagaa agttgatatt caacctcaca 5580 agggctctca aagtataatc tttctatagc caactgctaa tgcaaattaa aacatatttc 5640 attttaacat gatttcaaaa tcagtttttc atactaccct ttgctggaag aaactaaaaa 5700 tatagcaaat gcagaaccac aaacaattcg aatggggtag aaacattgta aatatttact 5760 ctttgcaaac cctggtggta ttttattttg gcttcatttc aatcattgaa gtatattctt 5820 attggaaatg tacttttgga taagtagggc taagccagtt ggatctctgg ttgtctagtc 5880 attgtcataa gtaaacctag taaaaccttg ttctattttt caatcatcaa aaagtaatta 5940 taaatacgta ttacaaacaa gtggatgttt ttaatgacca attgagtaag aacatccctg 6000 tcttaactgg cctaaatttc ttctggtagt gtcagttcaa ctttcagaag tgccacttaa 6060 ggaagtttga tttttgtttt tgtaatgcac tgtttttaat ctctctctct tttttttttt 6120 ttttttggtt ttaaaagcac aatcactaaa ctttatttgt aaaccattgt aactattaac 6180 cttttttgtc ttattgaaaa aaaaaatgtt gagaagcgtt tttaacctgt tttgttaatg 6240 ctctatgttt gtatttggaa tatttgaata atgacagatg gtgaagtaac atgcatactt 6300 tattgtgggc catgaaccaa atggttctta cttttcctgg acttaaagaa aaaaagaggt 6360 ttaagtttgt tgtggccaat gtcgaaacct acaagatttc cttaaaatct ctaatagagg 6420 cattacttgc tttcaattga caaatgatgc cctctgacta gtagatttct atgatccttt 6480 tttgtcattt tatgaatatc attgatttta taattggtgc tatttgaaga aaaaaatgta 6540 catttattca tagatagata agtatcaggt ctgaccccag tggaaaacaa agccaaacaa 6600 aactaacca caaaaaaaaa ggctggtgtt caccaaaacc aaacttgttc atttagataa 6660 tttgaaaaag ttccatagaa aaggcgtgca gtactaaggg aacaatccat gtgattaatg 6720 ttttcattat gttcatgtaa gaagcccctt atttttagcc ataattttgc atactgaaaa 6780 tccaataatc agaaaagtaa ttttgtcaca ttatttatta aaaatgttct caaatacata 6840 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaaaaaaaaaaaaaaaaa 6895 <210> 11 <211> 7550 <212> DNA <213> Homo sapiens roundabout, axon guidance receptor, homolog 1 (Drosophila) (ROBO1) <400> 11 aattgagctg gagaggaggc agcgtgagag cagaaacttc agacgccgct gatccgggag 60 gagctggggt gagccgcggc ggccgtctct cccacccgca gcagcatcct ctctgccctt 120 ctctgccag ccggggagag ccgggagctg cctctttaca gcttccacga gccaggggtg 180 caggcagctg cccccaggaa gtttgggctt ctgcgtagtt taggggtgcc tgcgagcgcc 240 ccagagggcg aggggccgag ggcgatgttg ggcgccgcgc ggggctgggg gcgcccagaa 300 gcgtgcgag tgtccgcggt cctgctgctg tctccagtac cctccgcatc ccccaagtga 360 tgggaacaag ggcccgccca ggcagccgct gtcgccgcac cgccccctcg ctcgctctct 420 gcgcgcggag tcacccagtc acactcccgg caccccgagc ccttcctccg gagctgctgc 480 ttctactttg gctgctatcg ccgccgccgc gggtggcccg ctgctgactg ggctcgccgg 540 gagacggaga agcacttttt ggccctccct cagcagctct cacaccccaa ctttgccgcc 600 gccgccgcgc ctgccctcgc agcggcgctc ggccgcacat tgtgggggcg cacgccggga 660 ggctccgcaa gaccgtggag gcaggaaacg gcactactgc gcttctgcct cggctctttg 720 ttgttcgctt tggatggttc ttgaaagtgt ctgagcctcc tcggaaatcc tggggccgga 780 gaagacaaac cttggaattc ttcctctgca aaagtctctg agatactgac aagcgtccgg 840 aaaggtcgac gagtaattgc cctgaaaact cttggctaat tgacccacgt tgcttatatt 900 aagcctttgt gtgtggtgtg tggcttcata catttgggga ccctatttcc actccctcct 960 cttggcatga gactgtatac aggatccacc cgaggacaat gattgcggag cccgctcact 1020 tttacctgtt tggattaata tgtctctgtt caggctcccg tcttcgtcag gaagattttc 1080 cacctcgcat tgttgaacac ccttcagacc tgattgtctc aaaaggagaa cctgcaactt 1140 tgaactgcaa agctgaaggc cgccccacac ccactattga atggtacaaa gggggagaga 1200 gagtggagac agacaaagat gaccctcgct cacaccgaat gttgctgccg agtggatctt 1260 tatttttctt acgtatagta catggacgga aaagtagacc tgatgaagga gtctatgtct 1320 gtgtagcaag gaattacctt ggagaggctg tgagccacaa tgcatcgctg gaagtagcca 1380 tacttcggga tgacttcaga caaaaccctt cggatgtcat ggttgcagta ggagagcctg 1440 cagtaatgga atgccaacct ccacgaggcc atcctgagcc caccatttca tggaagaaag 1500 atggctctcc actggatgat aaagatgaaa gaataactat acgaggagga aagctcatga 1560 tcacttacac ccgtaaaagt gacgctggca aatatgtttg tgttggtacc aatatggttg 1620 gggaacgtga gagtgaagta gccgagctga ctgtcttaga gagaccatca tttgtgaaga 1680 gacccagtaa cttggcagta actgtggatg acagtgcaga atttaaatgt gaggcccgag 1740 gtgaccctgt acctacagta cgatggagga aagatgatgg agagctgccc aaatccagat 1800 atgaaatccg agatgatcat accttgaaaa ttaggaaggt gacagctggt gacatgggtt 1860 catacacttg tgttgcagaa aatatggtgg gcaaagctga agcatctgct actctgactg 1920 ttcaagttgg gtctgaacct ccacattttg ttgtgaaacc ccgtgaccag gttgttgctt 1980 tgggacggac tgtaactttt cagtgtgaag caaccggaaa tcctcaacca gctattttct 2040 ggaggagaga agggagtcag aatctacttt tctcatatca accaccacag tcatccagcc 2100 gattttcagt ctcccagact ggcgacctca caattactaa tgtccagcga tctgatgttg 2160 gttattacat ctgccagact ttaaatgttg ctggaagcat catcacaaag gcatatttgg 2220 aagttacaga tgtgattgca gatcggcctc ccccagttat tcgacaaggt cctgtgaatc 2280 agactgtagc cgtggatggc actttcgtcc tcagctgtgt ggccacaggc agtccagtgc 2340 ccaccattct gtggagaaag gatggagtcc tcgtttcaac ccaagactct cgaatcaaac 2400 agttggagaa tggagtactg cagatccgat atgctaagct gggtgatact ggtcggtaca 2460 cctgcattgc atcaaccccc agtggtgaag caacatggag tgcttacatt gaagttcaag 2520 aatttggagt tccagttcag cctccaagac ctactgaccc aaatttaatc cctagtgccc 2580 catcaaaacc tgaagtgaca gatgtcagca gaaatacagt cacattatcg tggcaaccaa 2640 atttgaattc aggagcaact ccaacatctt atattataga agccttcagc catgcatctg 2700 gtagcagctg gcagaccgta gcagagaatg tgaaaacaga aacatctgcc attaaaggac 2760 tcaaacctaa tgcaatttac cttttccttg tgagggcagc taatgcatat ggaattagtg 2820 atccaagcca aatatcagat ccagtgaaaa cacaagatgt cctaccaaca agtcaggggg 2880 tggaccacaa gcaggtccag agagagctgg gaaatgctgt tctgcacctc cacaacccca 2940 ccgtcctttc ttcctcttcc atcgaagtgc actggacagt agatcaacag tctcagtata 3000 tacaaggata taaaattctc tatcggccat ctggagccaa ccacggagaa tcagactggt 3060 tagtttttga agtgaggacg ccagccaaaa acagtgtggt aatccctgat ctcagaaagg 3120 gagtcaacta tgaaattaag gctcgccctt tttttaatga atttcaagga gcagatagtg 3180 aaatcaagtt tgccaaaacc ctggaagaag cacccagtgc cccaccccaa ggtgtaactg 3240 tatccaagaa tgatggaaac ggaactgcaa ttctagttag ttggcagcca cctccagaag 3300 acactcaaaa tggaatggtc caagagtata aggtttggtg tctgggcaat gaaactcgat 3360 accacatcaa caaaacagtg gatggttcca ccttttccgt ggtcattccc tttcttgttc 3420 ctggaatccg atacagtgtg gaagtggcag ccagcactgg ggctgggtct ggggtaaaga 3480 gtgagcctca gttcatccag ctggatgccc atggaaaccc tgtgtcacct gaggaccaag 3540 tcagcctcgc tcagcagatt tcagatgtgg tgaagcagcc ggccttcata gcaggtattg 3600 gagcagcctg ttggatcatc ctcatggtct tcagcatctg gctttatcga caccgcaaga 3660 agagaaacgg acttactagt acctacgcgg gtatcagaaa agtaacttac cagagaggag 3720 gcgaagctgt cagcagtgga gggaggcctg gacttctcaa catcagtgaa cctgccgcgc 3780 agccatggct ggcagacacg tggcctaata ctggcaacaa ccacaatgac tgctccatca 3840 gctgctgcac ggcaggcaat ggaaacagcg acagcaacct cactacctac agtcgcccag 3900 ctgattgtat agcaaattat aacaaccaac tggataacaa acaaacaaat ctgatgctcc 3960 ctgagtcaac tgtttatggt gatgtggacc ttagtaacaa aatcaatgag atgaaaacct 4020 tcaatagccc aaatctgaag gatgggcgtt ttgtcaatcc atcagggcag cctactcctt 4080 acgccaccac tcagctcatc cagtcaaacc tcagcaacaa catgaacaat ggcagcgggg 4140 actctggcga gaagcactgg aaaccactgg gacagcagaa acaagaagtg gcaccagttc 4200 agtacaacat cgtggagcaa aacaagctga acaaagatta tcgagcaaat gacacagttc 4260 ctccaactat cccatacaac caatcatacg accagaacac aggaggatcc tacaacagct 4320 cagaccgggg cagtagtaca tctgggagtc aggggcacaa gaaaggggca agaacaccca 4380 aggtaccaaa acagggtggc atgaactggg cagacctgct tcctcctccc ccagcacatc 4440 ctcctccaca cagcaatagc gaagagtaca acatttctgt agatgaaagc tatgaccaag 4500 aaatgccatg tcccgtgcca ccagcaagga tgtatttgca acaagatgaa ttagaagagg 4560 aggaagatga acgaggcccc actccccctg ttcggggagc agcttcttct ccagctgccg 4620 tgtcctatag ccatcagtcc actgccactc tgactccctc cccacaggaa gaactccagc 4680 ccatgttaca ggattgtcca gaggagactg gccacatgca gcaccagccc gacaggagac 4740 ggcagcctgt gagtcctcct ccaccaccac ggccgatctc ccctccacat acctatggct 4800 acatttcagg acccctggtc tcagatatgg atacggatgc gccagaagag gaagaagacg 4860 aagccgacat ggaggtagcc aagatgcaaa ccagaaggct tttgttacgt gggcttgagc 4920 agacacctgc ctccagtgtt ggggacctgg agagctctgt cacggggtcc atgatcaacg 4980 gctggggctc agcctcagag gaggacaaca tttccagcgg acgctccagt gttagttctt 5040 cggacggctc ctttttcact gatgctgact ttgcccaggc agtcgcagca gcggcagagt 5100 atgctggtct gaaagtagca cgacggcaaa tgcaggatgc tgctggccgt cgacattttc 5160 atgcgtctca gtgccctagg cccacaagtc ccgtgtctac agacagcaac atgagtgccg 5220 ccgtaatgca gaaaaccaga ccagccaaga aactgaaaca ccagccagga catctgcgca 5280 gagaaaccta cacagatgat cttccaccac ctcctgtgcc gccacctgct ataaagtcac 5340 ctactgccca atccaagaca cagctggaag tacgacctgt agtggtgcca aaactccctt 5400 ctatggatgc aagaacagac agatcatcag acagaaaagg aagcagttac aagggagag 5460 aagtgttgga tggaagacag gttgttgaca tgcgaacaaa tccaggtgat cccagagaag 5520 cacaggaaca gcaaaatgac gggaaaggac gtggaaacaa ggcagcaaaa cgagaccttc 5580 caccagcaaa gactcatctc atccaagagg atattctacc ttattgtaga cctacttttc 5640 caacatcaaa taatcccaga gatcccagtt cctcaagctc aatgtcatca agaggatcag 5700 gaagcagaca aagagaacaa gcaaatgtag gtcgaagaaa tattgcagaa atgcaggtac 5760 ttggaggata tgaaagagga gaagataata atgaagaatt agaggaaact gaaagctgaa 5820 gacaaccaag aggcttatga gatctaatgt gaaaatcatc actcaagatg cctcctgtca 5880 gatgacacat gacgccagat aaaatgttca gtgcaatcag agtgtacaaa ttgtcgtttt 5940 tattcctctt attgggatat cattttaaaa actttattgg gtttttattg ttgttgtttg 6000 atccctaacc ctacaaagag ccttcctatt cccctcgctg ttggagcaaa ccattatacc 6060 ttacttccag caagcaaagt gctttgactt cttgcttcag tcatcagcca gcaagaggga 6120 acaaaactgt tcttttgcat tttgccgctg agatatggca ttgcactgct tatatgccaa 6180 gctaatttat agcaagatat tgatcaaata tagaaagttg atattcaacc tcacaagggc 6240 tctcaaagta taatctttct atagccaact gctaatgcaa attaaaacat atttcatttt 6300 aacatgattt caaaatcagt ttttcatact accctttgct ggaagaaact aaaaatatag 6360 caaatgcaga accacaaaca attcgaatgg ggtagaaaca ttgtaaatat ttactctttg 6420 caaaccctgg tggtatttta ttttggcttc atttcaatca ttgaagtata ttcttattgg 6480 aaatgtactt ttggataagt agggctaagc cagttggatc tctggttgtc tagtcattgt 6540 cataagtaaa cctagtaaaa ccttgttcta tttttcaatc atcaaaaagt aattataaat 6600 acgtattaca aacaagtgga tgtttttaat gaccaattga gtaagaacat ccctgtctta 6660 actggcctaa atttcttctg gtagtgtcag ttcaactttc agaagtgcca cttaaggaag 6720 tttgattttt gtttttgtaa tgcactgttt ttaatctctc tctctttttt tttttttttt 6780 tggttttaaa agcacaatca ctaaacttta tttgtaaacc attgtaacta ttaacctttt 6840 ttgtcttatt gaaaaaaaaa atgttgagaa gcgtttttaa cctgttttgt taatgctcta 6900 tgtttgtatt tggaatattt gaataatgac agatggtgaa gtaacatgca tactttattg 6960 tgggccatga accaaatggt tcttactttt cctggactta aagaaaaaaa gaggtttaag 7020 tttgttgtgg ccaatgtcga aacctacaag atttccttaa aatctctaat agaggcatta 7080 cttgctttca attgacaaat gatgccctct gactagtaga tttctatgat ccttttttgt 7140 cattttatga atatcattga ttttataatt ggtgctattt gaagaaaaaa atgtacattt 7200 attcatagat agataagtat caggtctgac cccagtggaa aacaaagcca aacaaaactg 7260 aaccacaaaa aaaaaggctg gtgttcacca aaaccaaact tgttcattta gataatttga 7320 aaaagttcca tagaaaaggc gtgcagtact aagggaacaa tccatgtgat taatgttttc 7380 attatgttca tgtaagaagc cccttatttt tagccataat tttgcatact gaaaatccaa 7440 taatcagaaa agtaattttg tcacattatt tattaaaaat gttctcaaat acataaaaaa 7500 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7550 <210> 12 <211> 7385 <212> DNA <213> Homo sapiens roundabout, axon guidance receptor, homolog 1 (Drosophila) (ROBO1) <400> 12 aattgagctg gagaggaggc agcgtgagag cagaaacttc agacgccgct gatccgggag 60 gagctggggt gagccgcggc ggccgtctct cccacccgca gcagcatcct ctctgccctt 120 ctctgccag ccggggagag ccgggagctg cctctttaca gcttccacga gccaggggtg 180 caggcagctg cccccaggaa gtttgggctt ctgcgtagtt taggggtgcc tgcgagcgcc 240 ccagagggcg aggggccgag ggcgatgttg ggcgccgcgc ggggctgggg gcgcccagaa 300 gcgtgcgag tgtccgcggt cctgctgctg tctccagtac cctccgcatc ccccaagtga 360 tgggaacaag ggcccgccca ggcagccgct gtcgccgcac cgccccctcg ctcgctctct 420 gcgcgcggag tcacccagtc acactcccgg caccccgagc ccttcctccg gagctgctgc 480 ttctactttg gctgctatcg ccgccgccgc gggtggcccg ctgctgactg ggctcgccgg 540 gagacggaga agcacttttt ggccctccct cagcagctct cacaccccaa ctttgccgcc 600 gccgccgcgc ctgccctcgc agcggcgctc ggccgcacat tgtgggggcg cacgccggga 660 ggctccgcaa gaccgtggag gcaggaaacg gcactactgc gcttctgcct cggctctttg 720 ttgttcgctt tggatggttc ttgaaagtgt ctgagcctcc tcggaaatcc tggggccgga 780 gaagacaaac cttggaattc ttcctctgca aaagtctctg agatactgac aagcgtccgg 840 aaaggtcgac gagtaattgc cctgaaaact cttggctaat tgacccacgt tgcttatatt 900 aagcctttgt gtgtggtgtg tggcttcata catttgggga ccctatttcc actccctcct 960 cttggcatga gactgtatac aggatccacc cgaggacaat gattgcggag cccgctcact 1020 tttacctgtt tggattaata tgtctctgtt caggctcccg tcttcgtcag gaagattttc 1080 cacctcgcat tgttgaacac ccttcagacc tgattgtctc aaaaggagaa cctgcaactt 1140 tgaactgcaa agctgaaggc cgccccacac ccactattga atggtacaaa gggggagaga 1200 gagtggagac agacaaagat gaccctcgct cacaccgaat gttgctgccg agtggatctt 1260 tatttttctt acgtatagta catggacgga aaagtagacc tgatgaagga gtctatgtct 1320 gtgtagcaag gaattacctt ggagaggctg tgagccacaa tgcatcgctg gaagtagcca 1380 tacttcggga tgacttcaga caaaaccctt cggatgtcat ggttgcagta ggagagcctg 1440 cagtaatgga atgccaacct ccacgaggcc atcctgagcc caccatttca tggaagaaag 1500 atggctctcc actggatgat aaagatgaaa gaataactat acgaggagga aagctcatga 1560 tcacttacac ccgtaaaagt gacgctggca aatatgtttg tgttggtacc aatatggttg 1620 gggaacgtga gagtgaagta gccgagctga ctgtcttaga gagaccatca tttgtgaaga 1680 gacccagtaa cttggcagta actgtggatg acagtgcaga atttaaatgt gaggcccgag 1740 gtgaccctgt acctacagta cgatggagga aagatgatgg agagctgccc aaatccagat 1800 atgaaatccg agatgatcat accttgaaaa ttaggaaggt gacagctggt gacatgggtt 1860 catacacttg tgttgcagaa aatatggtgg gcaaagctga agcatctgct actctgactg 1920 ttcaagttgg gtctgaacct ccacattttg ttgtgaaacc ccgtgaccag gttgttgctt 1980 tgggacggac tgtaactttt cagtgtgaag caaccggaaa tcctcaacca gctattttct 2040 ggaggagaga agggagtcag aatctacttt tctcatatca accaccacag tcatccagcc 2100 gattttcagt ctcccagact ggcgacctca caattactaa tgtccagcga tctgatgttg 2160 gttattacat ctgccagact ttaaatgttg ctggaagcat catcacaaag gcatatttgg 2220 aagttacaga tgtgattgca gatcggcctc ccccagttat tcgacaaggt cctgtgaatc 2280 agactgtagc cgtggatggc actttcgtcc tcagctgtgt ggccacaggc agtccagtgc 2340 ccaccattct gtggagaaag gatggagtcc tcgtttcaac ccaagactct cgaatcaaac 2400 agttggagaa tggagtactg cagatccgat atgctaagct gggtgatact ggtcggtaca 2460 cctgcattgc atcaaccccc agtggtgaag caacatggag tgcttacatt gaagttcaag 2520 aatttggagt tccagttcag cctccaagac ctactgaccc aaatttaatc cctagtgccc 2580 catcaaaacc tgaagtgaca gatgtcagca gaaatacagt cacattatcg tggcaaccaa 2640 atttgaattc aggagcaact ccaacatctt atattataga agccttcagc catgcatctg 2700 gtagcagctg gcagaccgta gcagagaatg tgaaaacaga aacatctgcc attaaaggac 2760 tcaaacctaa tgcaatttac cttttccttg tgagggcagc taatgcatat ggaattagtg 2820 atccaagcca aatatcagat ccagtgaaaa cacaagatgt cctaccaaca agtcaggggg 2880 tggaccacaa gcaggtccag agagagctgg gaaatgctgt tctgcacctc cacaacccca 2940 ccgtcctttc ttcctcttcc atcgaagtgc actggacagt agatcaacag tctcagtata 3000 tacaaggata taaaattctc tatcggccat ctggagccaa ccacggagaa tcagactggt 3060 tagtttttga agtgaggacg ccagccaaaa acagtgtggt aatccctgat ctcagaaagg 3120 gagtcaacta tgaaattaag gctcgccctt tttttaatga atttcaagga gcagatagtg 3180 aaatcaagtt tgccaaaacc ctggaagaag cacccagtgc cccaccccaa ggtgtaactg 3240 tatccaagaa tgatggaaac ggaactgcaa ttctagttag ttggcagcca cctccagaag 3300 acactcaaaa tggaatggtc caagagtata aggtttggtg tctgggcaat gaaactcgat 3360 accacatcaa caaaacagtg gatggttcca ccttttccgt ggtcattccc tttcttgttc 3420 ctggaatccg atacagtgtg gaagtggcag ccagcactgg ggctgggtct ggggtaaaga 3480 gtgagcctca gttcatccag ctggatgccc atggaaaccc tgtgtcacct gaggaccaag 3540 tcagcctcgc tcagcagatt tcagatgtgg tgaagcagcc ggccttcata gcaggtattg 3600 gagcagcctg ttggatcatc ctcatggtct tcagcatctg gctttatcga caccgcaaga 3660 agagaaacgg acttactagt acctacgcgg gtatcagaaa agtaacttac cagagaggag 3720 gcgaagctgt cagcagtgga gggaggcctg gacttctcaa catcagtgaa cctgccgcgc 3780 agccatggct ggcagacacg tggcctaata ctggcaacaa ccacaatgac tgctccatca 3840 gctgctgcac ggcaggcaat ggaaacagcg acagcaacct cactacctac agtcgcccag 3900 ggcagcctac tccttacgcc accactcagc tcatccagtc aaacctcagc aacaacatga 3960 acaatggcag cggggactct ggcgagaagc actggaaacc actgggacag cagaaacaag 4020 aagtggcacc agttcagtac aacatcgtgg agcaaaacaa gctgaacaaa gattatcgag 4080 caaatgacac agttcctcca actatcccat acaaccaatc atacgaccag aacacaggag 4140 gatcctacaa cagctcagac cggggcagta gtacatctgg gagtcagggg cacaagaaag 4200 gggcaagaac acccaaggta ccaaaacagg gtggcatgaa ctgggcagac ctgcttcctc 4260 ctcccccagc acatcctcct ccacacagca atagcgaaga gtacaacatt tctgtagatg 4320 aaagctatga ccaagaaatg ccatgtcccg tgccaccagc aaggatgtat ttgcaacaag 4380 atgaattaga agaggaggaa gatgaacgag gccccactcc ccctgttcgg ggagcagctt 4440 cttctccagc tgccgtgtcc tatagccatc agtccactgc cactctgact ccctccccac 4500 aggaagaact ccagcccatg ttacaggatt gtccagagga gactggccac atgcagcacc 4560 agcccgacag gagacggcag cctgtgagtc ctcctccacc accacggccg atctcccctc 4620 cacataccta tggctacatt tcaggacccc tggtctcaga tatggatacg gatgcgccag 4680 aagaggaaga agacgaagcc gacatggagg tagccaagat gcaaaccaga aggcttttgt 4740 tacgtgggct tgagcagaca cctgcctcca gtgttgggga cctggagagc tctgtcacgg 4800 ggtccatgat caacggctgg ggctcagcct cagaggagga caacatttcc agcggacgct 4860 ccagtgttag ttcttcggac ggctcctttt tcactgatgc tgactttgcc caggcagtcg 4920 cagcagcggc agagtatgct ggtctgaaag tagcacgacg gcaaatgcag gatgctgctg 4980 gccgtcgaca ttttcatgcg tctcagtgcc ctaggcccac aagtcccgtg tctacagaca 5040 gcaacatgag tgccgccgta atgcagaaaa ccagaccagc caagaaactg aaacaccagc 5100 caggacatct gcgcagagaa acctacacag atgatcttcc accacctcct gtgccgccac 5160 ctgctataaa gtcacctact gcccaatcca agacacagct ggaagtacga cctgtagtgg 5220 tgccaaaact cccttctatg gatgcaagaa cagacagatc atcagacaga aaaggaagca 5280 gttacaaggg gagagaagtg ttggatggaa gacaggttgt tgacatgcga acaaatccag 5340 gtgatcccag agaagcacag gaacagcaaa atgacgggaa aggacgtgga aacaaggcag 5400 caaaacgaga ccttccacca gcaaagactc atctcatcca agaggatatt ctaccttatt 5460 gtagacctac ttttccaaca tcaaataatc ccagagatcc cagttcctca agctcaatgt 5520 catcaagagg atcaggaagc agacaaagag aacaagcaaa tgtaggtcga agaaatattg 5580 cagaaatgca ggtacttgga ggatatgaaa gaggagaaga taataatgaa gaattagagg 5640 aaactgaaag ctgaagacaa ccaagaggct tatgagatct aatgtgaaaa tcatcactca 5700 agatgcctcc tgtcagatga cacatgacgc cagataaaat gttcagtgca atcagagtgt 5760 acaaattgtc gtttttattc ctcttattgg gatatcattt taaaaacttt attgggtttt 5820 tattgttgtt gtttgatccc taaccctaca aagagccttc ctattcccct cgctgttgga 5880 gcaaaccatt ataccttact tccagcaagc aaagtgcttt gacttcttgc ttcagtcatc 5940 agccagcaag agggaacaaa actgttcttt tgcattttgc cgctgagata tggcattgca 6000 ctgcttatat gccaagctaa tttatagcaa gatattgatc aaatatagaa agttgatatt 6060 caacctcaca agggctctca aagtataatc tttctatagc caactgctaa tgcaaattaa 6120 aacatatttc attttaacat gatttcaaaa tcagtttttc atactaccct ttgctggaag 6180 aaactaaaaa tatagcaaat gcagaaccac aaacattgg aatacgggtag aaacattgta 6240 aatatttact ctttgcaaac cctggtggta ttttattttg gcttcatttc aatcattgaa 6300 gtatattctt attggaaatg tacttttgga taagtagggc taagccagtt ggatctctgg 6360 ttgtctagtc attgtcataa gtaaacctag taaaaccttg ttctattttt caatcatcaa 6420 aaagtaatta taaatacgta ttacaaacaa gtggatgttt ttaatgacca attgagtaag 6480 aacatccctg tcttaactgg cctaaatttc ttctggtagt gtcagttcaa ctttcagaag 6540 tgccacttaa ggaagtttga tttttgtttt tgtaatgcac tgtttttaat ctctctctct 6600 tttttttttt ttttttggtt ttaaaagcac aatcactaaa ctttatttgt aaaccattgt 6660 aactattaac cttttttgtc ttattgaaaa aaaaaatgtt gagaagcgtt tttaacctgt 6720 tttgttaatg ctctatgttt gtatttggaa tatttgaata atgacagatg gtgaagtaac 6780 atgcatactt tattgtgggc catgaaccaa atggttctta cttttcctgg acttaaagaa 6840 aaaaagaggt ttaagtttgt tgtggccaat gtcgaaacct acaagatttc cttaaaatct 6900 ctaatagagg cattacttgc tttcaattga caaatgatgc cctctgacta gtagatttct 6960 atgatccttt tttgtcattt tatgaatatc attgatttta taattggtgc tatttgaaga 7020 aaaaaatgta catttattca tagatagata agtatcaggt ctgaccccag tggaaaacaa 7080 agccaaacaa aactgaacca caaaaaaaaa ggctggtgtt caccaaaacc aaacttgttc 7140 atttagataa tttgaaaaag ttccatagaa aaggcgtgca gtactaaggg aacaatccat 7200 gtgattaatg ttttcattat gttcatgtaa gaagcccctt atttttagcc ataattttgc 7260 atactgaaaa tccaataatc agaaaagtaa ttttgtcaca ttatttatta aaaatgttct 7320 caaatacata aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7380 aaaaa 7385

Claims

(a) a pharmaceutically effective amount of human SLIT2 (Slit homolog 2) protein or gene as an active ingredient; And (b) a pharmaceutically acceptable carrier. A pharmaceutical composition for the prevention or inhibition of lymph node metastasis or distant metastasis of primary thyroid cancer.

The composition of claim 1, wherein the thyroid cancer is thyroid papillary cancer.

delete

2. The composition of claim 1, wherein the composition reduces the rate of lymph node metastasis or the rate of metastasis of thyroid cancer.

The composition of claim 1, wherein the composition inhibits beta-catenin expression and induces E-cadherin activity in primary thyroid cancer cells.

2. The composition of claim 1, wherein said SLIT2 protein comprises the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3.