DOI QR코드

DOI QR Code

Molecular Cloning and Alternative Splicing of Growth Hormone Transcripts in Greenling, Hexagrammos otakii

쥐노래미 (Hexagrammos otakii) 성장호르몬 cDNA유전자의 염기서열 변이 및 발현 특성

  • Nam Yoon Kwon (Fish Cenetic Manipulation Laboratory, Department of Aquaculture, Pukyong National University) ;
  • Kim Dong Soo (Fish Cenetic Manipulation Laboratory, Department of Aquaculture, Pukyong National University)
  • Published : 2002.11.01

Abstract

Different types of transcripts encoding growth hormone (GH) were identified from cDNA libraries constructed with pituitaries of a marine fish species, greenling (Hexagrammos otakii). GH-homologous cDNA clones were isolated using the high-density filter hybridization and the expressed sequence tag techniques. Of 39 full-length positive cDNA clones, 31 clones ($79\%$) displayed an identical sequence, however, remaining 8 clones exhibited several polymorphisms in their sequences including (1) the length and sequence variability in the 5' upstream region, (2) insertional sequences in open reading frame, and (3) deletion and/or single nucleotide polymorphism in the untranslated 3' region. Based on RT-PCT and RNA dot blot analyses, these transcripts were proven to be expressed in a pituitary-specific manner.

우리나라 주요 해산 어종인 쥐노래미 (Hexagrammos otakii)로부터 성장호르몬 유전자 CDNA를 클로닝하고 이의 염기서열과 발현 특성을 분석하였다. 뇌하수체 조직으로부터 CDNA library를 제작하였으며 membrane filter hybridization 및 expressed sequence tag기술을 이용하여 성장호르몬 CDNA transcript들을 대량 발굴하였다. 총 확보된 full-length clone 39개중 31개가 동일한 형태로 나타났으나 나머지 클론들에서는 5'쪽의 염기서열 변이, ORF내의 염기서열 삽입, 3'쪽의 여기서열의 변이 등이 검출되었다. RT-PCR과 RNA dot blot 분석을 수행한 결과 본 연구에서 얻어진 쥐노래미 성장호르몬 transcript들은 뇌하수체 특이적인 전형적인 어류 성장호르몬 발현 특성을 나타내었다.

Keywords

References

  1. Agellon, L.B. and T.T. Chen. 1986. Rainbow trout growth hormone: molecular cloning of cDNA and expression in Escherichia coli. DNA, 5, 463-471 https://doi.org/10.1089/dna.1.1986.5.463
  2. Ber, R. and V. Daniel. 1993. Sequence analysis suggests a recent duplication of the growth honnone-encoding gene in Tilapia nilotica. Gene, 125, 143-150 https://doi.org/10.1016/0378-1119(93)90321-S
  3. Brett, D., J. Hanke, G. Lehmann, S. Haase, S. Delbruck, J. Reich and P. Bork. 2000. EST comparison indicates 38% of human mRNA contain possible alternative splice forms. FEBS Lett., 474, 83-86 https://doi.org/10.1016/S0014-5793(00)01581-7
  4. Chiou, C.-S., H.-T. Chen and W.-C. Chang. 1990. The complete nucleotide sequence of the growth-hormone gene from the common carp (Cyprinus carpio). Biochim. Biophy. Acta, 1087, 91-94 https://doi.org/10.1016/0167-4781(90)90126-M
  5. Devlin, R.H., T.Y. Yesaki, E.M. Donaldson, S.J. Du and C.L. Hew. 1995. Production of germline transgenic Pacific salmonids with dramatically increased growth performance. Can. J. Fish. Aquat. Sci., 52, 1376-1384 https://doi.org/10.1139/f95-133
  6. Douglas, S.E., J.W. Gallant, C.E. Bullerwell, C. Wolff, J. Munholland and M.E. Reith. 1999. Winter flounder expressed sequence tags: establishment of an EST database and identification of novel fish genes. Mar. Biotechnol., 1, 458-464 https://doi.org/10.1007/PL00011802
  7. Dunham, R.A. and R.H. Devlin. 1999. Comparison of traditional breeding and transgenesis in farmed fish with implications for growth enhancement and fitness. In: Transgenic Animals in Agriculture: Murray J.D., G.B. Anderson, A.M. Oberbauer and M.N. McGloughlin (eds). pp. 209-229. CAB International, Wallingford, UK
  8. Forbes, S.H., K.L. Knudsen, T.W. North and F.W. Allendorf. 1994. One of two growth hormone genes in coho salmon is sex-linked. Proc. Natl. Acad. Sci. USA, 91, 1628-1631 https://doi.org/10.1073/pnas.91.5.1628
  9. Gallo, A., E. Thomson, J. Brindle, M.A. O'Connell and L.P. Keegan. 2002. Micro precessing events in mRNAs identified by DHPLC analysis. Nucleic Acids Res., 30, 3945-3953 https://doi.org/10.1093/nar/gkf516
  10. Graveley, B.R. 2001. Alternative splicing: increasing diversity in the proteomic world. Trends Genet., 17, 100-107 https://doi.org/10.1016/S0168-9525(00)02176-4
  11. Hinits, Y. and B. Moav. 1999. Growth performance studies in transgenic Cyprinus carpio. Aquaculture, 173, 285-296 https://doi.org/10.1016/S0044-8486(98)00452-9
  12. Nam, Y.K. and D.S. Kim. 2001. Bioinformatics in fish: its present status and perspectives with particular emphasis on expressed sequence tags - brief review. J. Aquacult, 14, 9-16
  13. Nam, Y.K., J.K. Noh, Y.S. Cho, H.J. Cho, K.N. Cho, C.G. Kim and D.S. Kim. 2001. Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach (Misgurnus mizolepis). Transgenic Res., 10, 353-362 https://doi.org/10.1023/A:1016696104185
  14. Nam, Y.K., Y.S. Cho, S.E. Douglas, J.W. Gallant, M.E. Reith and D.S. Kim. 2002. Isolation and transient expression of a cDNA encoding L-gulono-g-lactone oxidase, a key enzyme for L-ascorbic acid biosynthesis, from the tiger shark Scyliorhinus torazame. Aquaculture, 209, 271-284 https://doi.org/10.1016/S0044-8486(01)00731-1
  15. Shen, H., H. Illges, A. Reuter and C.A.O. Stuenner. 2002. Cloning, expression, and alternative splicing of neogenin 1 in zebrafish. Mech. Dev., 118, 219-223 https://doi.org/10.1016/S0925-4773(02)00255-1
  16. Young, G., B.T. Bjornsson, P. Prunet, R.J. Lin and H.A. Bern. 1989. Smoltification and seawater adaptation in coho salmon (Oncorhynchus kisutch): plasma prolactin, growth hormone, thyroid hormones, and cortisol. Gene. Comp. Endocrinol., 74, 335-345 https://doi.org/10.1016/S0016-6480(89)80029-2
  17. Untergasser, G., M. Hermann, H. Rumpold, G. Pfister and P. Berger. 2000. An unusual member of the human growth hormone/placental lactogen (GH/PL) family, the testicular altemative splicing variant hPL-A2: recombinant expression revealed a membrane-associated growth factor molecule. Mol. Cell. Endocnnol., 167, 117-125 https://doi.org/10.1016/S0303-7207(00)00287-2