CN1182245C - Brachydactyly and body height associated gene - Google Patents

Abstract

The present invention relates to a brachydactyly and body height associated gene, the coding of which has a protein A-1 type polypeptide with amino acid sequence of SEQ ID NO. 2. The brachydactyly and body height associated gene has nucleotide sequence of SEQ ID NO. 1 and comprises a gene IHH in the section of 2q35 to q36. The coding of a gene IHH in the region of Exon1 has a protein polypeptide with amino acid sequence of SEQ ID NO. 6. The present invention clones an A-1 type brachydactyly and body height associated gene, namely that the present invention positions the gene IHH in the section of 2q35 to q36 and determines a new mutant site of the gene, so that the present invention provides a reference of gene relation for molecular genetics interrelation for A-1 type brachydactyly pathogenesis and body height development. The present invention effectively promotes the further development of human bone dysplasia pathomechanism research, deepens human knowledge on bone development at a molecular level and provides genetics references for corresponding gene diagnosis, drug design and clinical treatment for the future.

Description

Brachydactylia and height-related gene

Technical field: the present invention relates to a kind of brachydactylia and height-related gene, relate more specifically to the sequence of a kind of A-1 type brachydactylia and height-related gene and this dna encoding the protein molecule.The gene field that belongs to biotechnology.

(brachydactyly, BD) be divided into five types is A, B, C, D and E to background technology: Bell (Treasury of Human inheritance.1951.5:1-30) with the heredity brachydactylia.Bell has found three hypotype A-1, A-2 and the A-3 hypotype in the A type.A-1 type brachydactylia (BDA-1, principal character MIM:112500) is the shortening of middle dactylus, middle dactylus that has and far-end dactylus merge or middle dactylus disappearance.A-1 type brachydactylia be eighties of last century by the human Mendelian autosomal dominant inherited disease of Farabeo (1903) reported first, and list in the textbook as exemplary, it is typical single-gene dominant hereditary disease.(Mendelian inheritence in man.Johns Hopkins University Press.1975 Baltimore) has increased A-4 and A-5 type brachydactylia to Mckusick.The the 2nd and the 5th middle dactylus that refers to shortens the 4th dactylus bending in A-4 type brachydactylia.In A-5 type brachydactylia, all middle dactylus all lack.Fitch (J MedGenet.1979.16 (1): 36-44) reexamined the X-ray sheet of 44 A-1 types, the A-1 type has been done systematic research. find because each shortening that refers to, whole palm seems short and small, and the hand bone is also short than normal hand bone, shortens the most serious with middle dactylus especially.Therefore, Fitch also belongs to the A-1 type with A-4 and A-5 type brachydactylia.The A-1 type is sometimes with other complication, as the women in a family stiff and dysnoesia (the Piussan et al.J Genet Hum.1983.31 (2): 107-114) of thumb joint is arranged, and an A-1 type patient who has 5q11.2 and 17q23 balanced translocation has unusual (the Slavatinek etal.Clin Dysmorphol.1998.7 (1): 21-27) of Klippel-Feil (neck throat heteroplasia).The adjusting of IHH signal path ginseng skeletal growth, growth, its mutant can cause the shortening of experiment mice bone to be proved (Chua ng et al, Nature, 1999,397:617-621; St-Jacques et al., Genes ﹠amp; Development, 1999,13:2072-2086).

The applicant has formally proposed " a kind of Short finger gene " January 4 calendar year 2001 to Patent Office of the People's Republic of China, and application number is 01105018.7, has described in this patent application to have cloned an A-1 type brachydactylia refer to ospc gene in two big A-1 type brachydactylia familys.

Summary of the invention: the clinical symptom of the 3rd the A-1 type brachydactylia family that the present invention obtains with original have obviously different, be in particular in: the patient has the different middle dactylus of degree to lose or significantly shortens hypertrophy with fusion symptom, the thumb of other dactylus, the more important thing is that patient's height (between 150 to 160 centimetres) significantly is lower than the height (between 165 to 180 centimetres) of normal individual in the family.By to corresponding gene clone, order-checking research, found a new gene mutation site, this sudden change is not only relevant with A-1 type brachydactylia, and relevant with the growth of height.For this reason, content of the present invention relates to a kind of A-1 type brachydactylia and height-related gene sequence and homologous variation gene order thereof of isolated or purifying, the sequence that also relates to the said gene encode protein molecule, said " isolated or purifying " be meant (so-called isolated or purified is meant on the Genomic dna level, utilize primer for the amplification of each exon and utilize purification kit that amplified production is purified).A-1 type brachydactylia and height-related gene sequence are shown in SEQ NO.1:

(a) sequence signature:

^*Length: 1236 base pairs

^*Type: nucleic acid

^*Chain: two strands

^*Topological framework: linear

(b) molecule type: mRNA (cDNA library)

(c) suppose: not

(d) antisense: not

(e) initial source: people

(f) SEQ ID NO.1 nucleotide sequence is described below:

ATGTCTCCCGCCCGGCTCCGGCCCCGACTGCACTTCTGCCTGGTCCTGTT

GCTGCTGCTGGTGGTGCCGGCGGCATGGGGCTGCGGGCCGGGTCGGGTG

GTGGGCAGCCGCCGGCGACCGCCACGCAAACTCGTGCCGCTCGCCTACA

AGCAGTTCAGCCCCAATGTGCCCGAGAAGACCCTGGGCGCCAGCGGACG

CTATGAAGGCAAGATCGCTCGCAGCTCCGAGCGCTTCAAGGAGCTCACC

CCCAATTACAATCCAGACATCATCTTCAAGGACGAGGAGAACACAGGCG

CCGACCGCCTCATGACCCAGCGCTGCAAGGACCGCCTGAACTCGCTGGC

TATCTCGGTGATGAACCAGTGGCCCGGTGTGAAGCTGCGGGTGACCGAG

GGCTGGGACGAGGACGGCCACCACTCAGAGGAGTCCCTGCATTATGAGG

GCCGCGCGGTGGACATCACCACATCAGACCGCGACCGCAATAAGTATGG

ACTGCTGGCGCGCTTGGCAGTGGAGGCCGGCTTTGACTGGGTGTATTAC

GAGTCAAAGGCCCACGTGCATTGCTCCGTCAAGTCCGAGCACTCGGCCG

CAGCAACGACGGGCGGCTGCTTCCCTGCCGGAGCCCAGGTACGCCTGGA

GAGTGGGGCGCGTGTGGCCTTGTCAGCCGTGAGGCCGGGAGACCGTGTG

CTGGCCATGGGGGAGGATGGGAGCCCCACCTTCAGCGATGTGCTCATTTT

CCTGGACCGCGAGCCTCACAGGCTGAGAGCCTTCCAGGTCATCGAGACT

CAGGACCCCCCACGCCGCCTGGCACTCACACCCGCTCACCTGCTCTTTAC

GGCTGACAATCACACGGAGCCGGCAGCCCGCTTCCGGGCCACATTTGCC

AGCCACGTGCAGCCTGGCCAGTACGTGCTGGTGGCTGGGGTGCCAGGCC

TGCAGCCTGCCCGCGTGGCAGCTGTCTCTACACACGTGGCCCTCGGGGC

CTACGCCCCGCTCACAAAGCATGGGACACTGGTGGTGGAGGATGTGGTG

GCATCCTGCTTCGCGGCCGTGGCTGACCACCACCTGGCTCAGTTGGCCTT

CTGGCCCCTGAGACTCTTTCACAGCTTGGCATGGGGCAGCTGGACCCCG

GGGGAGGGTGTGCATTGGTACCCCCAGCTGCTCTACCGCCTGGGGCGTC

TCCTGCTAGAAGAGGGCAGCTTCCACCCACTGGGCATGTCCGGGGCAGG

GAGCTGA

That base position that the representative of the letter of dash area is undergone mutation in the sequence.

The sequence of said gene encode protein molecule is shown in SEQ NO.2:

(a) sequence signature:

^*Length: 411 amino acid

^*Type: amino acid

^*Chain: strand

^*Topological framework: linear

(b) molecule type: peptide

(c) SEQ ID NO.2 aminoacid sequence is described below:

1?msparlrprl?hfclvlllll?vvpaawgcgp?grvvgsrrrp?prklvplayk?qfspnvpekt

61?lgasgryegk?iarsserfke?ltpnynpdii?fkdeentga rlmtqrckdr?lnslaisvmn

121?qwpgvklrvt?egwdedghhs?eeslhyegra?vdittsdrdr?nkygllarla?veagfdwvyy

181?eskahvhcsv?ksehsaaakt?ggcfpagaqv?rlesgarval?savrpgdrvl?amgedgsptf

241?sdviilldre?phrlrafqfi?etqdpprrla?ltpahllfta?dnhtepaarf?ratfashvqp

301?gqyvlvagvp?glqparvaav?sthvalgaya?pltkhgtlvv?edvvascfaa?vadhhlaqla

361?fwplrlfhsl?awgswtpgeg?vhwypqllyr?lgrllleegs?fhplgmsgag?s

That amino acid position that the representative of the letter of dash area is undergone mutation in the sequence.

Above-mentioned A-1 type Short finger gene sequence SEQ NO.1 has a kind of varient, and its coding has the homologous variation sequence that is less than 8 sequence changes, and the change of sequence is the change of conservative property sequence, and its gene order is shown in SEQ ID NO.3:

(a) sequence signature:

^*Length: 1236 base pairs

^*Type: nucleic acid

^*Chain: two strands

^*Topological framework: linear

(b) molecule type: mRNA (cDNA library)

(c) suppose: not

(d) antisense: not

(e) initial source: people

(f) SEQ ID NO.3 nucleotide sequence is described below:

ATGTCTCCCGCCCGGCTCCGGCCCCGACTGCACTTCTGCCTGGTCCTGTT

GCTGCTGCTGGTGGTGCCGGCGGCATGGGGCTGCGGGCCGGGTCGGGTG

GTGGGCAGCCGCCGGCGACCGCCACGCAAACTCGTGCCGCTCGCCTACA

AGCAGTTCAGCCCCAATGTGCCCGAGAAGACCCTGGGCGCCAGCGGACG

CTATGAAGGCAAGATCGCTCGCAGCTCCGAGCGCTTCAAGGAGCTCACC

CCCAATTACAATCCAGACATCATCTTCAAGGACGAGGAGAACACAGGCG

CCGA CGCCTCATGACCCAGCGCTGCAAGGACCGCCTGAACTCGCTGGC

TATCTCGGTGATGAACCAGTGGCCCGGTGTGAAGCTGCGGGTGACCGAG

GGCTGGGACGAGGACGGCCACCACTCAGAGGAGTCCCTGCATTATGAGG

GCCGCGCGGTGGACATCACCACATCAGACCGCGACCGCAATAAGTATGG

ACTGCTGGCGCGCTTGGCAGTGGAGGCCGGCTTTGACTGGGTGTATTAC

GAGTCAAAGGCCCACGTGCATTGCTCCGTCAAGTCCGAGCACTCGGCCG

CAGCAACGACGGGCGGCTGCTTCCCTGCCGGAGCCCAGGTACGCCTGGA

GAGTGGGGCGCGTGTGGCCTTGTCAGCCGTGAGGCCGGGAGACCGTGTG

CTGGCCATGGGGGAGGATGGGAGCCCCACCTTCAGCGATGTGCTCATTTT

CCTGGACCGCGAGCCTCACAGGCTGAGAGCCTTCCAGGTCATCGAGACT

CAGGACCCCCCACGCCGCCTGGCACTCACACCCGCTCACCTGCTCTTTAC

GGCTGACAATCACACGGAGCCGGCAGCCCGCTTCCGGGCCACATTTGCC

AGCCACGTGCAGCCTGGCCAGTACGTGCTGGTGGCTGGGGTGCCAGGCC

TGCAGCCTGCCCGCGTGGCAGCTGTCTCTACACACGTGGCCCTCGGGGC

CTACGCCCCGCTCACAAAGCATGGGACACTGGTGGTGGAGGATGTGGTG

GCATCCTGCTTCGCGGCCGTGGCTGACCACCACCTGGCTCAGTTGGCCTT

CTGGCCCCTGAGACTCTTTCACAGCTTGGCATGGGGCAGCTGGACCCCG

GGGGAGGGTGTGCATTGGTACCCCCAGCTGCTCTACCGCCTGGGGCGTC

TCCTGCTAGAAGAGGGCAGCTTCCACCCACTGGGCATGTCCGGGGCAGG

GAGCTGA

That base position that the representative of the letter of dash area is undergone mutation in the sequence.

The varient of above-mentioned gene order, its coding has the homologous variation albumen that is less than 8 amino acid changes, and amino acid change is that conservative amino acid changes, and it has the sequence shown in the SEQ ID NO.4:

(a) sequence signature:

^*Length: 411 amino acid

^*Type: amino acid

^*Chain: strand

^*Topological framework: linear

(b) molecule type: peptide

(c) SEQ ID NO.4 aminoacid sequence is described below:

1?msparlrprl?hfclvlllll?vvpaawgcgp?grvvgsrrrp?prklvplayk?qfspnvpekt

61?lgasgryegk?iarsserfke?ltpnynpdii?fkdeentga rlmtqrckdr?lnslaisvmn

121?qwpgvklrvt?egwdedghhs?eeslhyegra?vdittsdrdr?nkygllarla?veagfdwvyy

181?eskahvhcsv?ksehsaaakt?ggcfpagaqv?rlesgarval?savrpgdrvl?amgedgsptf

241?sdviilldre?phrlrafqfi?etqdpprrla?ltpahllfta?dnhtepaarf?ratfashvqp

301?gqyvlvagvp?glqparvaav?sthvalgaya?pltkhgtlvv?edvvascfaa?vadhhlaqla

361?fwplrlfhsl?awgswtpgeg?vhwypqllyr?lgrllleegs?fhplgmsgag?s

That amino acid position that the representative of the letter of dash area is undergone mutation in the sequence.

The mRNA sequence in the Exon1 zone of normal human subject IHH is an Exon1 part among the SEQ ID NO.1, and both comprise and involved relation, and its sequence is shown in SEQ ID NO.5:

SEQ ID NO.5 nucleotide sequence is described below:

Exon1 (normal people)

ATGTCTCCCGCCCGGCTCCGGCCCCGACTGCACTTCTGCCTGGTCCTGTT

GCTGCTGCTGGTGGTGCCGGCGGCATGGGGCTGCGGGCCGGGTCGGGTG

GTGGGCAGCCGCCGGCGACCGCCACGCAAACTCGTGCCGCTCGCCTACA

AGCAGTTCAGCCCCAATGTGCCCGAGAAGACCCTGGGCGCCAGCGGACG

CTATGAAGGCAAGATCGCTCGCAGCTCCGAGCGCTTCAAGGAGCTCACC

CCCAATTACAATCCAGACATCATCTTCAAGGACGAGGAGAACACAGGCG

CCGA

CGCCTCATGACCCAG

That base position that the representative of the letter of dash area is undergone mutation in the sequence.

The protein polypeptide that the described genes encoding of sequence SEQ ID NO.5 has SEQ ID NO.6 sequence, SEQ ID NO.6 aminoacid sequence is described below:

Exon1 proteins encoded (normal people)

MSPARLRPRLHFCLVLLLLLVVPAAWGCGPGRVVGSRRRPPRKLVPLAYKQF

SPNVPEKTLGASGRYEGKIARSSERFKELTPNYNPDIIFKDEENTGA RLMT

Q

That amino acid position that the representative of the letter of dash area is undergone mutation in the sequence.

SEQ ID NO.5 has a homologous variation body, i.e. the mRNA sequence in the Exon1 zone of patient's IHH, i.e. and SEQ ID NO.7, SEQ ID NO.7 nucleotide sequence is described below:

Exon1 (patient)

ATGTCTCCCGCCCGGCTCCGGCCCCGACTGCACTTCTGCCTGGTCCTGTT

GCTGCTGCTGGTGGTGCCGGCGGCATGGGGCTGCGGGCCGGGTCGGGTG

GTGGGCAGCCGCCGGCGACCGCCACGCAAACTCGTGCCGCTCGCCTACA

AGCAGTTCAGCCCCAATGTGCCCGAGAAGACCCTGGGCGCCAGCGGACG

CTATGAAGGCAAGATCGCTCGCAGCTCCGAGCGCTTCAAGGAGCTCACC

CCCAATTACAATCCAGACATCATCTTCAAGGACGAGGAGAACACAGGCG

CCGA CGCCTCATGACCCAG

That base position that the representative of the letter of dash area is undergone mutation in the sequence.

Accordingly, the protein polypeptide that above-mentioned mRNA sequence SEQ ID NO.7 coding has SEQ ID NO.8 sequence, i.e. SEQ ID NO.8, SEQ ID NO.8 aminoacid sequence is described below:

Exon1 proteins encoded (patient)

MSPARLRPRLHFCLVLLLLLVVPAAWGCGPGRVVGSRRRPPRKLVPLAYKQF

SPNVPEKTLGASGRYEGKIARSSERFKELTPNYNPDIIFKDEENTGA RLMTQ

That amino acid position that the representative of the letter of dash area is undergone mutation in the sequence.

Sequence SEQ ID NO.5-SEQ ID NO.8 infers like this:

Can obtain following data through the GeneBank search database:

A.L38517?Homo?sapiens?indian?hedgehog?protein(IHH)mRNA，5′end

B.AB021874?Homo?sapiens?hedgehog?gene，5′-flanking?region，partialcds

C.U85610?Mus?musculus?Indian?hedgehog?protein(Ihh)mRNA，completecds

D.Q14623?INDIAN?HEDGEHOG?PROTEIN?PRECURSOR(IHH)(HHG-2)

C is that the full length mRNA of mouse is divided into three exons, Exon1, Exon2, Exon3. but in the database only human IHH data be A.B.Wherein A only comprises Exon2 and the Exon3 homologous sequence of C, there is no Exon1, and B then is 5 ' non-coding sequence.The full length mRNA sequence of no human IHH in the database.Homology is very high as can be known by analyzing mouse and human existing sequence, utilize the human Genomic database of Exon1 sequence search of mouse can obtain the zone of a matched, by analyzing this zone, obtained a kind of mutant, this zone is the Exon1 zone of human IHH, obtain the full length mRNA of human IHH by splicing, translate into find behind the albumen with database in existing human IHH protein D. coincide.

The present invention will produce very important and far-reaching influence in the genetic field for diagnosis, treatment and the height growth of the relevant genetic diseases of bone, at first, the present invention has cloned a kind of A-1 type brachydactylia and height-related gene promptly is positioned 2q35-q36 section IHH gene, determine the new mutational site of this gene, and finished the research of structure and function; Secondly, the present invention provides the foundation of gene relationship for the mutual relationship of the molecular genetic of morbidity of A-1 type brachydactylia and height growth, this invention will effectively promote the further research of human skeletal's heteroplasia pathomechanism, deepen mankind's understanding to skeleton development on molecular level, and for corresponding gene diagnosis, medicinal design and clinical treatment provide the basis of genetics from now on.

Description of drawings:

Fig. 1 is the clinical photo of BDA-1

Fig. 2 is the PCR product glue figure of Exon1

Fig. 3 is an A-1 type gene PCR amplified production sequence chart

Fig. 4 is the AciI restriction enzyme mapping

Fig. 5 is the variation of wild-type and the coded proteinic three-dimensional space conformation of mutated genes

Embodiment: collect the 3rd A-1 family totally 5 patients from the remote mountain areas in Hunan Province, utilize in these two big familys of X-ray examination patient to go smoothly and foot, the data that provides meets the description of Fitch about non-syndrome type BDA-1 feature.As seen from Figure 1, as Fig. 1-a, A-1 type brachydactylia characteristic feature refers to (toe) joint disappearance in the middle of being, or refers to that with the centre (toe) joint and far-end merge; As Fig. 1-c, patient (among the figure placed in the middle and occupy right person) height is starkly lower than the normal people's (occupying left person among the figure) in the family.

On the basis of the above, utilize conventional pcr amplification candidate dna and dna sequencing analysis, clone, enzyme technology such as to cut, A-1 type brachydactylia and height-related gene have been cloned, being positioned 2q35-q36 section IHH gene is exactly A-1 type brachydactylia and height-related gene, and has found a new mutational site.Just because of the IHH gene this sudden change having taken place, has caused the on the low side of the morbidity of A-1 type brachydactylia and patient's height.Sudden change occurs in the coding region of Exon1 (No. 1 exon on) of this family IHH gene, and has obtained checking on all patients' the DNA sample in family.By pcr amplification, product purification, the method for order-checking obtains the Exon1 sequence, concrete grammar step following (wherein listed Exon1 method is the amplification that comprises the sudden change part among the Exon1, does not comprise total length):

Exon1：

PCR reaction system: (the every reagent of system provides by Bio-Asia company except that DNA, and the back together)

DdH ₂The O 16.5ul distilled water of sterilizing

Buffer 2.5ul polyreaction damping fluid comprises KCl Tris-HCl solution

Mg ²⁺2.0ul 25mM MgCl ₂Solution

DNTP 2.0ul 2mM dNTP solution

Pri-F 0.4ul 20pmol/ul forward primer 5 '-CGGACGCTATGAAGGCAAGA-3 '

Pri-R 0.4ul 20pmol/ul reverse primer 5 '-GCCAGCCAGTCGAGAAAATG-3 '

Taq 0.2ul DNA synthesizes polymerase

DNA 1.0ul 20ng/ul human DNA amplification template

Volume?25.0ul

The PCR reaction conditions:

1、95℃ 5’

2、95℃ 30”

3、58℃?45”

4、72℃?1’

5、Go?to?step?2?for?35?cycles

6、72℃?10’

7、4℃ hold

1.5％agarose?gel?check

The PCR product glue figure of Exon1 sees Fig. 2, and the figure left side is the 100bp trapezoidal shape.

96 orifice plates are all adopted in above PCR reaction, and MJ Research PTC-225 Peltier Thermal CyclerPCR instrument carries out, and glue figure photo adopts the picked-up of the Bio-Rad Gel-Doc of company video camera.

The order-checking of A-1 type gene PCR amplified production:

The PCR product adopts Promega Wizard Kit purifying, by specification operation

The sequencing reaction system:

DdH ₂The O 2ul distilled water of sterilizing

DNA behind the DNA 5ul purifying, meter 200ng

Buffer 2.5ul order-checking buffer (autogamy 1M Tric-Hcl: 25mM Mgcl ₂: ddH ₂O 2: 2: 1)

BDT 2.5ul ABI?BigDyeTerminator?Cylce?sequencing?Ready?Reaction?Kit

The 3pmol/ul of primer forward or backwards of the PCR product that Primer 0.8ul is obtained

The sequencing reaction condition:

1、96℃ 2’

2、96℃ 10”

3、50℃ 5”

4、60℃ 4’

5、Go?to?step?2?for?40?cycles

6、4℃ hold

Finish the back and add 70% ethanol lucifuge and left standstill 15 minutes, 4000 rev/mins centrifugal 30 minutes, get rid of ethanol, essence is put and was dried in 1 hour, adds sample in the loading buffer preparation.Order-checking adopts ABI PRISM377 DNA Sequencer to carry out.

Sequence chart is seen Fig. 3, and Fig. 3-a is the normal people, and Fig. 3-b is patient.Utilize the enzyme blanking method that A-1 gene PCR amplified production is verified that the restriction analysis method is as follows simultaneously:

The Exon1 product adopts the TaKaRa AciI of biotech company digestion with restriction enzyme

AciI 0.5ul(5U) TaKaRa?Biotechnology?Co.ltd

Buffer?2.5ul TaKaRa?Biotechnology?Co.ltd

DNA 10ul PCR product

DdH ₂The O 7ul distilled water of sterilizing

volume?20ul

37 ℃ 8 hours

Adopt 20% polyacrylamide gel to detect, the AciI restriction enzyme mapping is seen Fig. 4, the left side is the PCR product, and cut for normal people's enzyme the centre, and cut for patient's enzyme on the right, and change has also taken place in wild-type and the coded proteinic three-dimensional space conformation of mutated genes, the results are shown in Figure 5, Fig. 5-a is a normal circumstances, and 100As shown by arrows, Fig. 5-b is the situation that 100Asp sports 100Glu, and Fig. 5-c is the secondary structure model diagram of regular.Particularly, be that the 300th C (base) on the Exon1 sports A (base) in the family in this Hunan Province, make the 100th Asp (amino acid) in the wild-type Exon1 encoded protein matter molecule change Glu (amino acid) (base mutation is seen Fig. 3, and amino acid mutation is seen Fig. 5) into.Have following biological activity based on wild-type Exon1 encoded protein matter: in the IHH signal pathway that the limb development process is played a crucial role, the proteic N-terminal of IHH is a signal domain, is exactly the coded zone of Exon1.Infer the coded protein in wild-type Exon1 sudden change back thus, because the change that amino acid is formed, thereby reduce or changed the biological activity that this protein is just often had, hindered the normal growth and the growth of osseous tissue, cause the generation of A-1 type and the reduction of height.

In the specification sheets about nucleotide sequence and aminoacid sequence data

SEQ ID NO.1 nucleotide sequence is described below:

ATGTCTCCCGCCCGGCTCCGGCCCCGACTGCACTTCTGCCTGGTCCTGTT

GCTGCTGCTGGTGGTGCCGGCGGCATGGGGCTGCGGGCCGGGTCGGGTG

GTGGGCAGCCGCCGGCGACCGCCACGCAAACTCGTGCCGCTCGCCTACA

AGCAGTTCAGCCCCAATGTGCCCGAGAAGACCCTGGGCGCCAGCGGACG

CTATGAAGGCAAGATCGCTCGCAGCTCCGAGCGCTTCAAGGAGCTCACC

CCCAATTACAATCCAGACATCATCTTCAAGGACGAGGAGAACACAGGCG

CCGACCGCCTCATGACCCAGCGCTGCAAGGACCGCCTGAACTCGCTGGC

TATCTCGGTGATGAACCAGTGGCCCGGTGTGAAGCTGCGGGTGACCGAG

GGCTGGGACGAGGACGGCCACCACTCAGAGGAGTCCCTGCATTATGAGG

GCCGCGCGGTGGACATCACCACATCAGACCGCGACCGCAATAAGTATGG

ACTGCTGGCGCGCTTGGCAGTGGAGGCCGGCTTTGACTGGGTGTATTAC

GAGTCAAAGGCCCACGTGCATTGCTCCGTCAAGTCCGAGCACTCGGCCG

CAGCAACGACGGGCGGCTGCTTCCCTGCCGGAGCCCAGGTACGCCTGGA

GAGTGGGGCGCGTGTGGCCTTGTCAGCCGTGAGGCCGGGAGACCGTGTG

CTGGCCATGGGGGAGGATGGGAGCCCCACCTTCAGCGATGTGCTCATTTT

CCTGGACCGCGAGCCTCACAGGCTGAGAGCCTTCCAGGTCATCGAGACT

CAGGACCCCCCACGCCGCCTGGCACTCACACCCGCTCACCTGCTCTTTAC

GGCTGACAATCACACGGAGCCGGCAGCCCGCTTCCGGGCCACATTTGCC

AGCCACGTGCAGCCTGGCCAGTACGTGCTGGTGGCTGGGGTGCCAGGCC

TGCAGCCTGCCCGCGTGGCAGCTGTCTCTACACACGTGGCCCTCGGGGC

CTACGCCCCGCTCACAAAGCATGGGACACTGGTGGTGGAGGATGTGGTG

GCATCCTGCTTCGCGGCCGTGGCTGACCACCACCTGGCTCAGTTGGCCTT

CTGGCCCCTGAGACTCTTTCACAGCTTGGCATGGGGCAGCTGGACCCCG

GGGGAGGGTGTGCATTGGTACCCCCAGCTGCTCTACCGCCTGGGGCGTC

TCCTGCTAGAAGAGGGCAGCTTCCACCCACTGGGCATGTCCGGGGCAGG

GAGCTGA

That base position that the representative of the letter of dash area is undergone mutation in the sequence.

SEQ ID NO.2 aminoacid sequence is described below:

1?msparlrprl?hfclvlllll?vvpaawgcgp?grvvgsrrrp?prklvplayk?qfspnvpekt

61?lgasgryegk?iarsserfke?ltpnynpdii?fkdeentgad?rlmtqrckdr?lnslaisvmn

121?qwpgvklrvt?egwdedghhs?eeslhyegra?vdittsdrdr?nkygllarla?veagfdwvyy

181?eskahvhcsv?ksehsaaakt?ggcfpagaqv?rlesgarval?savrpgdrvl?amgedgsptf

241?sdviilldre?phrlrafqfi?etqdpprrla?ltpahllfta?dnhtepaarf?ratfashvqp

301?gqyvlvagvp?glqparvaav?sthvalgaya?pltkhgtlvv?edvvascfaa?vadhhlaqla

361?fwplrlfhsl?awgswtpgeg?vhwypqllyr?lgrllleegs?fhplgmsgag?s

That amino acid position that the representative of the letter of dash area is undergone mutation in the sequence.

SEQ ID NO.3 nucleotide sequence is described below:

ATGTCTCCCGCCCGGCTCCGGCCCCGACTGCACTTCTGCCTGGTCCTGTT

GCTGCTGCTGGTGGTGCCGGCGGCATGGGGCTGCGGGCCGGGTCGGGTG

GTGGGCAGCCGCCGGCGACCGCCACGCAAACTCGTGCCGCTCGCCTACA

AGCAGTTCAGCCCCAATGTGCCCGAGAAGACCCTGGGCGCCAGCGGACG

CTATGAAGGCAAGATCGCTCGCAGCTCCGAGCGCTTCAAGGAGCTCACC

CCCAATTACAATCCAGACATCATCTTCAAGGACGAGGAGAACACAGGCG

CCGAACGCCTCATGACCCAGCGCTGCAAGGACCGCCTGAACTCGCTGGC

TATCTCGGTGATGAACCAGTGGCCCGGTGTGAAGCTGCGGGTGACCGAG

GGCTGGGACGAGGACGGCCACCACTCAGAGGAGTCCCTGCATTATGAGG

GCCGCGCGGTGGACATCACCACATCAGACCGCGACCGCAATAAGTATGG

ACTGCTGGCGCGCTTGGCAGTGGAGGCCGGCTTTGACTGGGTGTATTAC

GAGTCAAAGGCCCACGTGCATTGCTCCGTCAAGTCCGAGCACTCGGCCG

CAGCAACGACGGGCGGCTGCTTCCCTGCCGGAGCCCAGGTACGCCTGGA

GAGTGGGGCGCGTGTGGCCTTGTCAGCCGTGAGGCCGGGAGACCGTGTG

CTGGCCATGGGGGAGGATGGGAGCCCCACCTTCAGCGATGTGCTCATTTT

CCTGGACCGCGAGCCTCACAGGCTGAGAGCCTTCCAGGTCATCGAGACT

CAGGACCCCCCACGCCGCCTGGCACTCACACCCGCTCACCTGCTCTTTAC

GGCTGACAATCACACGGAGCCGGCAGCCCGCTTCCGGGCCACATTTGCC

AGCCACGTGCAGCCTGGCCAGTACGTGCTGGTGGCTGGGGTGCCAGGCC

TGCAGCCTGCCCGCGTGGCAGCTGTCTCTACACACGTGGCCCTCGGGGC

CTACGCCCCGCTCACAAAGCATGGGACACTGGTGGTGGAGGATGTGGTG

GCATCCTGCTTCGCGGCCGTGGCTGACCACCACCTGGCTCAGTTGGCCTT

CTGGCCCCTGAGACTCTTTCACAGCTTGGCATGGGGCAGCTGGACCCCG

GGGGAGGGTGTGCATTGGTACCCCCAGCTGCTCTACCGCCTGGGGCGTC

TCCTGCTAGAAGAGGGCAGCTTCCACCCACTGGGCATGTCCGGGGCAGG

GAGCTGA

That base position that the representative of the letter of dash area is undergone mutation in the sequence.SEQ ID NO.4 aminoacid sequence is described below:

1?msparlrprl?hfclvlllll?vvpaawgcgp?grvvgsrrrp?prklvplayk?qfspnvpekt

61?lgasgryegk?iarsserfke?ltpnynpdii?fkdeentgac?rlmtqrckdr?lnslaisvmn

121?qwpgvklrvt?egwdedghhs?eeslhyegra?vdittsdrdr?nkygllarla?veagfdwvyy

181?eskahvhcsv?ksehsaaakt?ggcfpagaqv?rlesgarval?savrpgdrvl?amgedgsptf

241?sdviilldre?phrlrafqfi?etqdpprrla?ltpahllfta?dnhtepaarf?ratfashvqp

301?gqyvlvagvp?glqparvaav?sthvalgaya?pltkhgtlvv?edvvascfaa?vadhhlaqla

361?fwplrlfhsl?awgswtpgeg?vhwypqllyr?lgrllleegs?fhplgmsgag?s

That amino acid position that the representative of the letter of dash area is undergone mutation in the sequence.

SEQ ID NO.5 nucleotide sequence is described below:

Exon1 (normal people)

ATGTCTCCCGCCCGGCTCCGGCCCCGACTGCACTTCTGCCTGGTCCTGTT

GCTGCTGCTGGTGGTGCCGGCGGCATGGGGCTGCGGGCCGGGTCGGGTG

GTGGGCAGCCGCCGGCGACCGCCACGCAAACTCGTGCCGCTCGCCTACA

AGCAGTTCAGCCCCAATGTGCCCGAGAAGACCCTGGGCGCCAGCGGACG

CTATGAAGGCAAGATCGCTCGCAGCTCCGAGCGCTTCAAGGAGCTCACC

CCCAATTACAATCCAGACATCATCTTCAAGGACGAGGAGAACACAGGCG

CCGACCGCCTCATGACCCAG

That base position that the representative of the letter of dash area is undergone mutation in the sequence.

SEQ ID NO.6 preface amino acid row are described below:

Exon1 proteins encoded (normal people)

MSPARLRPRLHFCLVLLLLLVVPAAWGCGPGRVVGSRRRPPRKLVPLAYKQF

SPNVPEKTLGASGRYEGKIARSSERFKELTPNYNPDIIFKDEENTGADRLMT

Q

That amino acid position that the representative of the letter of dash area is undergone mutation in the sequence.

SEQ ID NO.7 nucleotide sequence is described below:

Exon1 (patient)

ATGTCTCCCGCCCGGCTCCGGCCCCGACTGCACTTCTGCCTGGTCCTGTT

GCTGCTGCTGGTGGTGCCGGCGGCATGGGGCTGCGGGCCGGGTCGGGTG

GTGGGCAGCCGCCGGCGACCGCCACGCAAACTCGTGCCGCTCGCCTACA

AGCAGTTCAGCCCCAATGTGCCCGAGAAGACCCTGGGCGCCAGCGGACG

CTATGAAGGCAAGATCGCTCGCAGCTCCGAGCGCTTCAAGGAGCTCACC

CCCAATTACAATCCAGACATCATCTTCAAGGACGAGGAGAACACAGGCG

CCGAACGCCTCATGACCCAG

That base position that the representative of the letter of dash area is undergone mutation in the sequence.

SEQ ID NO.8 aminoacid sequence is described below:

Exon1 proteins encoded (patient)

MSPARLRPRLHFCLVLLLLLVVPAAWGCGPGRVVGSRRRPPRKLVPLAYKQF

SPNVPEKTLGASGRYEGKIARSSERFKELTPNYNPDIIFKDEENTGAERLMTQ

That amino acid position that the representative of the letter of dash area is undergone mutation in the sequence.

Claims (2)

1. brachydactylia and height-related gene is characterized in that its nucleotide sequence is shown in SEQ ID NO.3.

2. the varient of brachydactylia as claimed in claim 2 and height-related gene is characterized in that, it has the aminoacid sequence shown in the SEQ ID NO.4.

Images