JPS63216899A - Growth hormone 1002 of fishes - Google Patents
Growth hormone 1002 of fishesInfo
- Publication number
- JPS63216899A JPS63216899A JP62049999A JP4999987A JPS63216899A JP S63216899 A JPS63216899 A JP S63216899A JP 62049999 A JP62049999 A JP 62049999A JP 4999987 A JP4999987 A JP 4999987A JP S63216899 A JPS63216899 A JP S63216899A
- Authority
- JP
- Japan
- Prior art keywords
- growth hormone
- fish
- fishes
- growth
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000018997 Growth Hormone Human genes 0.000 title claims abstract description 34
- 108010051696 Growth Hormone Proteins 0.000 title claims abstract description 34
- 241000251468 Actinopterygii Species 0.000 title claims abstract description 22
- 239000000122 growth hormone Substances 0.000 title claims abstract description 15
- 235000019688 fish Nutrition 0.000 title abstract description 16
- 241001441724 Tetraodontidae Species 0.000 claims abstract description 9
- 150000001413 amino acids Chemical class 0.000 claims abstract description 9
- 239000000203 mixture Substances 0.000 claims abstract description 9
- NPWMTBZSRRLQNJ-VKHMYHEASA-N (3s)-3-aminopiperidine-2,6-dione Chemical compound N[C@H]1CCC(=O)NC1=O NPWMTBZSRRLQNJ-VKHMYHEASA-N 0.000 claims abstract description 4
- 238000010521 absorption reaction Methods 0.000 claims abstract description 3
- 239000000126 substance Substances 0.000 claims abstract 2
- 238000000862 absorption spectrum Methods 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 239000007864 aqueous solution Substances 0.000 claims 1
- 229920001184 polypeptide Polymers 0.000 claims 1
- 102000004196 processed proteins & peptides Human genes 0.000 claims 1
- 108090000765 processed proteins & peptides Proteins 0.000 claims 1
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 6
- 206010062767 Hypophysitis Diseases 0.000 abstract description 5
- 210000003635 pituitary gland Anatomy 0.000 abstract description 5
- 230000001737 promoting effect Effects 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- 239000003795 chemical substances by application Substances 0.000 abstract 1
- 239000000727 fraction Substances 0.000 abstract 1
- 238000001641 gel filtration chromatography Methods 0.000 abstract 1
- 239000005556 hormone Substances 0.000 abstract 1
- 229940088597 hormone Drugs 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 abstract 1
- 241000277275 Oncorhynchus mykiss Species 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 8
- 230000037396 body weight Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000000872 buffer Substances 0.000 description 4
- 230000001900 immune effect Effects 0.000 description 4
- 230000001766 physiological effect Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 241001600434 Plectroglyphidodon lacrymatus Species 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 230000000366 juvenile effect Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000277329 Oncorhynchus keta Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 description 1
- 241000881711 Acipenser sturio Species 0.000 description 1
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 description 1
- 229910000013 Ammonium bicarbonate Inorganic materials 0.000 description 1
- 241000283153 Cetacea Species 0.000 description 1
- 241000252233 Cyprinus carpio Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102000015731 Peptide Hormones Human genes 0.000 description 1
- 108010038988 Peptide Hormones Proteins 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 241000609666 Tuber aestivum Species 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 235000012538 ammonium bicarbonate Nutrition 0.000 description 1
- 239000001099 ammonium carbonate Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000009260 cross reactivity Effects 0.000 description 1
- 125000001295 dansyl group Chemical group [H]C1=C([H])C(N(C([H])([H])[H])C([H])([H])[H])=C2C([H])=C([H])C([H])=C(C2=C1[H])S(*)(=O)=O 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 125000000404 glutamine group Chemical group N[C@@H](CCC(N)=O)C(=O)* 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 150000002825 nitriles Chemical class 0.000 description 1
- 239000000813 peptide hormone Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
Abstract
Description
【発明の詳細な説明】
本発明は、魚類の成長ホルモンポリペプチドに関し、詳
しくはフグ目、フグ科魚類の成長ホルモンポリペプチド
に関し、更に詳しくはフグ目フグ科フグ属魚類の成長ホ
ルモンポリペプチドに関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a growth hormone polypeptide of fish, particularly to a growth hormone polypeptide of fish of the order Pufferidae and family Pufferidae, and more particularly to a growth hormone polypeptide of fish of the order Pufferidae, family Pufferidae, genus Pufferfish. .
成長ホルモンは、を推動物の脳下垂体から分泌されるポ
リペプチドホルモンで、その主たる生理作用は成長促進
効果であることが知られている。Growth hormone is a polypeptide hormone secreted from the pituitary gland of growing animals, and its main physiological action is known to be a growth-promoting effect.
これまでに、哺乳類ではヒト、ヒツジ、ブタ、クジラ、
ウマなどで成長ホルモンポリペブチＶが単離され、構造
が決定されている。So far, mammals such as humans, sheep, pigs, whales,
The growth hormone polypeptide V has been isolated from horses and other species, and its structure has been determined.
一方、下等を推動物、特に魚類ではティラビア、チョウ
ザメ、コイ、シロザケおよびウナギで成長ホルモンポリ
ペプチドが単離され、このうち、シロザケ、ウナギにつ
いては最近その構造が明らかとなった。On the other hand, growth hormone polypeptides have been isolated from lower mammals, especially fish, such as tilavia, sturgeon, carp, chum salmon, and eel, and the structures of chum salmon and eel have recently been clarified.
これらの知見によると、同じ魚類でも科、属が異なれば
、成長ホルモンポリペプチドの構造は各々異なり、また
、生理活性（成長促進効果）においても種特異性がある
こと、即ち、成長ホルモンポリペプチドをそれを単離し
た魚種に投与する方が池の魚種に投与するより効果が著
しいことが示されている。According to these findings, the structures of growth hormone polypeptides differ among fish from different families and genera, and there is also species-specificity in physiological activity (growth-promoting effect). It has been shown that administering the drug to isolated fish species is more effective than administering it to pond fish species.
従って、特定の魚種の成長ホルモンポリペプチドを抽出
し同定することが出来れば、魚類の養殖産業分野におい
て幅広い用途が期待される。Therefore, if it is possible to extract and identify the growth hormone polypeptide of a specific fish species, a wide range of applications are expected in the fish aquaculture industry.
そこで、本発明者らは、日本の重要魚種で、かつ養殖技
術が確立されているフグから成長ホルモンポリペプチド
を単離し同定することが出来れば、この成長ホルモンポ
リペプチドをフグに投与することにより、現在約２年を
要する養殖期間を大幅に短縮させることが出来ると考え
、鋭意研究の結果、本発明に達するに至った。即ち、本
発明によれば、フグの脳下垂体より成長ホルモンポリペ
プチドを抽出し、精製して同定するとともに、これをニ
ジマスの稚魚に投与することにより、著しい成長促進効
果を確認することが出来たものである。Therefore, the present inventors believe that if it is possible to isolate and identify a growth hormone polypeptide from pufferfish, which is an important fish species in Japan and for which aquaculture technology has been established, it would be possible to administer this growth hormone polypeptide to pufferfish. We believed that this would greatly shorten the cultivation period, which currently takes about two years, and as a result of intensive research, we arrived at the present invention. That is, according to the present invention, by extracting, purifying and identifying growth hormone polypeptide from the pituitary gland of puffer fish, and administering it to juvenile rainbow trout, it was possible to confirm a remarkable growth-promoting effect. It is something that
以下、本発明によるフグの成長ホルモンポリペプチドの
単離と同定法についてその概要を述べる。Hereinafter, the method for isolating and identifying growth hormone polypeptide of puffer fish according to the present invention will be summarized.
フグ脳下垂体のアルカリ抽出物を順次デル濾過、高速液
体クロマトグラフィー（ＨＰＬＣ）を行うことにより、
純粋な成長ホルモンポリペプチドを単離、精製した。By sequentially performing del filtration and high performance liquid chromatography (HPLC) on an alkaline extract of pufferfish pituitary gland,
Pure growth hormone polypeptide was isolated and purified.
その結果、フグの成長ホルモンポリペプチドは、分子量
約２２，０００．構成アミノ酸の組成は、同じ海産魚の
ハマチと同様にグルタミン酸、ロイシンが多く認められ
、等電点は、約６．４〜６．５であった。また、本発明
の成長ホルモンポリペプチドの生理活性について、ニジ
マス稚魚を用いて、体長、体重の促進効果を調べた結果
、体長、体重とも著しい促進効果が認められた。また、
本発明の成長ホルモンポリペプチドは、同じ海産魚であ
るハマチの成長ホルモンの特異抗体とゲル内二重拡散法
で沈降線を得たことから、免疫学的にも成長ホルモンで
あることが実証された。As a result, the growth hormone polypeptide of puffer fish has a molecular weight of about 22,000. As for the constituent amino acid composition, glutamic acid and leucine were found in large amounts, similar to the same marine fish yellowtail, and the isoelectric point was about 6.4 to 6.5. Furthermore, regarding the physiological activity of the growth hormone polypeptide of the present invention, the effect of promoting body length and body weight was investigated using juvenile rainbow trout, and as a result, a remarkable promoting effect on both body length and body weight was observed. Also,
The growth hormone polypeptide of the present invention was immunologically proven to be a growth hormone, as a sedimentation line was obtained using the in-gel double diffusion method with a specific antibody for growth hormone from the same marine fish, yellowtail. Ta.
以下、実施例により本発明を更に詳細に説明する。Hereinafter, the present invention will be explained in more detail with reference to Examples.
ｍ工：成長ホルモンポリペプチドの抽出２年魚の生きて
いるフグから摘出し、すぐ液体窒素中で凍結させ、使用
時主で一８０°Ｃに保存した脳下垂体（約４００個、８
ｇ、）を氷冷中で摩砕後、１０（ｈｌの冷アセトンで
脱脂した。これを５ｍＨＥＤＴ＾および１．５ｍＨＰＭ
ＳＦ（酵素失活剤）を含む１００ａ＋１の氷冷水に懸濁
させ、水冷中で１時間抽出した。M Engineering: Extraction of Growth Hormone Polypeptide Pituitary glands (approximately 400 pieces, 8
g,) was ground in ice-cooling and degreased with 10 (hl) of cold acetone.
The suspension was suspended in 100a+1 ice-cold water containing SF (enzyme inactivator) and extracted for 1 hour under water cooling.
抽出液は一４℃、 １５，０００回転、３０分間遠心分
離し、その上清をＹＭ−２メンプランによる限外濾過で
濃縮後、デル濾過に添加した。The extract was centrifuged at -4° C. and 15,000 rpm for 30 minutes, and the supernatant was concentrated by ultrafiltration using YM-2 Membrane and then added to a Delfilter filter.
実施例２ ；成長ホルモンポリペプチドの精製（１）
ゲル濾過
抽出物を０．０５Ｍ重炭酸アンモニウム緩衝液（ｐＨ９
，０）で平衡化したセフ７デツクスＧ−７５（カラムサ
イズ： ２．６４Ｘ ６５．Ｏｃｍ）で分画した。但し
、溶出条件は前流を１００ｍＬ流速は２２．５＋Ｎ／時
、試験管１本あたりの分収量は３ｍＮとし、２８０ｎｍ
の吸光度でタンパク含量を測定した。その結果、第１図
に示したように、４つの区分に分画させた。尚、区分ご
との収量を第１表に示した。Example 2; Purification of growth hormone polypeptide (1)
The gel-filtered extract was dissolved in 0.05M ammonium bicarbonate buffer (pH 9).
Fractionation was carried out using Cef7dex G-75 (column size: 2.64×65.Ocm) equilibrated with 2.64×65.Ocm). However, the elution conditions are 100 mL of the front flow, a flow rate of 22.5+N/hour, a yield of 3 mN per test tube, and a 280 nm
The protein content was measured by the absorbance of . As a result, as shown in FIG. 1, it was fractionated into four sections. The yield for each category is shown in Table 1.
第１表
（２） 高速液体クロマトグラフィー（ＨＰＬＣ）デル
濾過で分画された４つの区分のうち、第３の区分をＯ［
１Ｓ−１２０Ｔカラム（カラムサイズ ：０．４Ｘ ２
４．Ｏｃｍ）を用いて逆相型のＨＰＬＣで精製した。Table 1 (2) Among the four fractions fractionated by high performance liquid chromatography (HPLC) del filtration, the third fraction was
1S-120T column (column size: 0.4X 2
4. It was purified by reverse-phase HPLC using HPLC (Ocm).
但し、溶出条件は０．１％トリフ０口酢酸、アセトニト
リル４０％〜１００％直線勾配、カラム温度４０℃、流
速６０ａ＋１／時とし、２２０ｎｍの吸光度チタンバク
含量を測定して、溶出液を各区分ごとに分取した。However, the elution conditions were 0.1% truffle 0-mouth acetic acid, 40% to 100% linear gradient of acetonitrile, column temperature 40°C, flow rate 60a+1/hour, and the absorbance content at 220 nm was measured, and the eluate was divided into each section. It was separated into
その結果、第２図に示したように、アセ）ニトリル濃度
７６％で溶出された１０番目の区分（７ラクシジンＡ）
（総収量２．ＱＢ）について以下に示すように物理化学
的特性、生理活性および免疫学的特性について検討した
。As a result, as shown in Figure 2, the 10th segment (7 Laccidine A) was eluted at an ace)nitrile concentration of 76%.
(total yield 2.QB) was examined for physicochemical properties, physiological activity, and immunological properties as shown below.
実施例３ ：物理化学的特性の測定
５ＯＳ−ゲル電気泳動を用いて分子量測定を行った結果
、約２２，０００であった。等電点測定には、ファルマ
シアのＰｈａｓｔ ｓｙｓｔｅｍの電気泳動装置とＰｈ
ａｓｔ ８ｅｌ（ｐｌ（３〜９ ）を用い、その結果、
約６．４〜６．５であった。また、紫外線吸収スペクト
ルにおける最大吸収は２１０ｎｍであった。Example 3: Measurement of physicochemical properties The molecular weight was measured using 5OS-gel electrophoresis, and the result was approximately 22,000. For isoelectric point measurement, Pharmacia's Phast system electrophoresis device and Ph
using ast 8el(pl(3-9), resulting in
It was about 6.4-6.5. Further, the maximum absorption in the ultraviolet absorption spectrum was 210 nm.
実施例・（ニアミノ酸組成の決定
６Ｎ−塩酸で減圧密封後、１１０℃、２４時間で加水分
解を行い、アミノ酸分析機を用いてアミノ酸の組成を決
定した。その結果を既知の魚類の成長ホルモンのアミノ
酸組成と共に第２表に示した。成長ホルモンのアミノ酸
組成の特徴は、グルタミン酸およびロイシンが多いこと
であるが、本発明の成長ホルモンポリペプチドにおいて
も同様な傾向が顕著に見られることがわかった。Example: (Determination of amino acid composition) After sealing under reduced pressure with 6N hydrochloric acid, hydrolysis was carried out at 110°C for 24 hours, and the composition of amino acids was determined using an amino acid analyzer.The results were compared with known fish growth hormones. The amino acid composition of growth hormone is shown in Table 2.The amino acid composition of growth hormone is characterized by a high content of glutamic acid and leucine, and it was found that the same tendency is clearly seen in the growth hormone polypeptide of the present invention. Ta.
第２表 アミノ酸組成１）
１） アミノ酸残基数はトリプトファンを１個とした時
の相対数を表す。Table 2 Amino acid composition 1) 1) The number of amino acid residues represents the relative number when one tryptophan is used.
実施例５ ニアミノ末端分析 アミノ末端分析はダンシル法に上り行った。Example 5 Niamino terminal analysis Amino terminal analysis was performed using the Dansyl method.
その結果、ダンシル−アミノ酸は検出されず、従って、
アミノ末端はグルタミンが環をまいているピログルタミ
ンであると推定された。As a result, dansyl-amino acids were not detected, and therefore,
The amino terminal was presumed to be pyroglutamine with a glutamine ring.
実施例６ ：生理活性の測定
実装点として約ｆｏｃｍ、約１０ｇのニジマス稚魚を用
い、本発明の成長ホルモンポリペプチドを生理食塩水に
溶解し、ニジマスの腹腔内に注射により７日おきに４回
投与した。１回の投与量は、ニジマスの体重８あたり１
μｇＩ Ｏ，１μ８およびコントロールとして生理食塩
水のみの計３群（１群１０匹）に分けて行い、餌は朝夕
２回、飽食まで与えた。各点の体長および体重を測定し
、伸長効果を下記の式に従って伸長率で表した結果を第
３図および第４図に示す。Example 6: Measurement of physiological activity Using approximately 10 g of juvenile rainbow trout as a mounting point, the growth hormone polypeptide of the present invention was dissolved in physiological saline and injected intraperitoneally into the rainbow trout 4 times every 7 days. administered. One dose is 1/8 of the body weight of rainbow trout.
The mice were divided into 3 groups (10 animals per group) containing only μgI O, 1 μ8 and physiological saline as a control, and were fed twice in the morning and once in the evening until satiated. The body length and weight at each point were measured, and the elongation effect was expressed as an elongation rate according to the following formula. The results are shown in FIGS. 3 and 4.
図から明らかなように、本発明の成長ホルモンポリペプ
チド１土、ニジマスに対して著しい成長促進効果が認め
られた。As is clear from the figure, the growth hormone polypeptide 1 of the present invention had a significant growth promoting effect on rainbow trout.
実施例７ ：免疫学的同定
成長ホルモンポリペプチドは、ハマチの成長ホルモンの
抗体とゲル内二重拡散法で免疫交差性がみちれた。よっ
て、免疫学的にも成長ホルモンであることが、実証され
た。Example 7: Immunological Identification The growth hormone polypeptide was found to have immunological cross-reactivity with the yellowtail growth hormone antibody using the in-gel double diffusion method. Therefore, it was demonstrated that it is a growth hormone immunologically as well.
以上の結果から、本発明の成長ホルモンポリペプチドは
既知の魚類の成長ホルモンと物理化学的性質および免疫
学的性質において一致し、アミノ酸１１成も類年１する
こと、また、生理活性の測定において、異種のニジマス
においてさえも著しい伸長効果を与えることからフグの
成長ホルモンと同定した。From the above results, the growth hormone polypeptide of the present invention is identical to known fish growth hormones in physicochemical and immunological properties, and the composition of 11 amino acids is similar to that of known fish growth hormones. It was identified as the growth hormone of pufferfish because it has a remarkable elongation effect even in different species of rainbow trout.
第１図は、実施例２の（１）ゲル濾過に述べたセファデ
ックスＧ−７５カラムでの溶出パターンを示す。
第２図は、実施例２の（２）高速液体クロマトグラフィ
ーに述べた高速液体クロマトグラフイー（ＯＤＳ−１２
０Ｔカラム）での溶出パターンを示す。
第３図および第４図は、ニジマスに本発明の成長ホルモ
ンポリペプチドを投与した後の日ごとの体長および体重
の伸長率を示す。
尚、図中のΔは対照群、○は体重ｇあたり０．１μｇ投
与群、および・は体重８あたり１．０μｇ投与群を示す
。
特許出願人 株式会社アドバンス
（他１名）
第１図
チェーフ’ｆ；Ａ−−
を−千２FIG. 1 shows the elution pattern on the Sephadex G-75 column described in Example 2 (1) Gel filtration. Figure 2 shows the high performance liquid chromatography (ODS-12) described in Example 2 (2) High performance liquid chromatography.
0T column) is shown. Figures 3 and 4 show the daily growth rate of body length and body weight after administering the growth hormone polypeptide of the present invention to rainbow trout. In the figure, Δ indicates a control group, ○ indicates a group administered with 0.1 μg per gram of body weight, and ◯ indicates a group administered with 1.0 μg per gram of body weight. Patent applicant Advance Co., Ltd. (1 other person) Figure 1 Chaef'f;A-- 1,000
Claims (2)
成長ホルモン。 （ｉ）アミノ酸組成 第２表に記載の通り。 （ｉｉ）分子量 約２２，０００ （ｉｉｉ）等電点 ６．４〜６．５ （ｉｖ）水溶液に可溶 （ｖ）アミノ末端アミノ酸 ピログルタミン （ｖｉ）紫外吸収スペクトル：最大吸収２１０ｎｍ(1) A polypeptide fish growth hormone having the following physical and chemical properties. (i) Amino acid composition As described in Table 2. (ii) Molecular weight: approximately 22,000 (iii) Isoelectric point: 6.4-6.5 (iv) Soluble in aqueous solution (v) Amino-terminal amino acid pyroglutamine (vi) Ultraviolet absorption spectrum: maximum absorption 210 nm
）項記載の成長ホルモン。(2) Claim No. 1 in which the fish belongs to the Pufferfish family, Pufferfish genus.
Growth hormones listed in ).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62049999A JPS63216899A (en) | 1987-03-06 | 1987-03-06 | Growth hormone 1002 of fishes |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62049999A JPS63216899A (en) | 1987-03-06 | 1987-03-06 | Growth hormone 1002 of fishes |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS63216899A true JPS63216899A (en) | 1988-09-09 |
Family
ID=12846703
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62049999A Pending JPS63216899A (en) | 1987-03-06 | 1987-03-06 | Growth hormone 1002 of fishes |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63216899A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110558256A (en) * | 2019-10-09 | 2019-12-13 | 中国水产科学研究院黄海水产研究所 | nutrition method for regulating and controlling bile acid secretion of takifugu rubripes in programmed mode |
-
1987
- 1987-03-06 JP JP62049999A patent/JPS63216899A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110558256A (en) * | 2019-10-09 | 2019-12-13 | 中国水产科学研究院黄海水产研究所 | nutrition method for regulating and controlling bile acid secretion of takifugu rubripes in programmed mode |
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