Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1981 Feb 25;9(4):935–946. doi: 10.1093/nar/9.4.935

Use of a cloned double stranded cDNA coding for a major androgen dependent protein in rat seminal vesicle secretion: the effect of testosterone in gene expression.

P E Mansson, A Sugino, S E Harris
PMCID: PMC326723  PMID: 6164983

Abstract

The abundant class of poly(A+)RNA [poly(A+)RNA11S] from rat seminal vesicle was used to synthesize ds-cDNA11S. The ds-cDNA11S was inserted and cloned into the Pst I site of pBR-322 using E. coli RR1 as host. Colony filter hybridization and restriction mapping was used to demonstrate that a 620 NTP long insert in a plasmid clone (pSV2) represents the almost full length structural gene coding for a precursor to the seminal vesicle secretion protein IV (SVS IV). The entire insert was sequenced and the coding region was matched with the known amino acid sequence. Most of the signal peptide sequence was derived from the DNA sequence. The insert in pSV2 was labelled and used to study the effect of testosterone on the accumulation of mRNA SVS IV. Administration of testosterone to castrated rats resulted in the induction of mRNA SVS IV from a few molecules per cell to levels of over 100,000 after 96 h of hormone treatment.

Full text

PDF
935

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Guyette W. A., Matusik R. J., Rosen J. M. Prolactin-mediated transcriptional and post-transcriptional control of casein gene expression. Cell. 1979 Aug;17(4):1013–1023. doi: 10.1016/0092-8674(79)90340-4. [DOI] [PubMed] [Google Scholar]
  3. Harris S. E., Rosen J. M., Means A. R., O'Malley B. W. Use of a specific probe for ovalbumin messenger RNA to quantitate estrogen-induced gene transcripts. Biochemistry. 1975 May 20;14(10):2072–2081. doi: 10.1021/bi00681a006. [DOI] [PubMed] [Google Scholar]
  4. Higgins S. J., Burchell J. M. Effects of testosterone on messenger ribonucleic acid and protein synthesis in rat seminal vesicle. Biochem J. 1978 Aug 15;174(2):543–551. doi: 10.1042/bj1740543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Higgins S. J., Burchell J. M., Mainwaring W. I. Androgen-dependent synthesis of basic secretory proteins by the rat seminal vesicle. Biochem J. 1976 Aug 15;158(2):271–282. doi: 10.1042/bj1580271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Higgins S. J., Burchell J. M., Mainwaring W. I. Testosterone control of nucleic acid content and proliferation of epithelium and stroma in rat seminal vesicles. Biochem J. 1976 Oct 15;160(1):43–48. doi: 10.1042/bj1600043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Higgins S. J., Parker M. G., Fuller F. M., Jackson P. J. Androgenic regulation of messenger RNA sequence complexity in accessory sexual tissues of the male rat studied with fractionated complementary DNA. Eur J Biochem. 1979 Dec 17;102(2):431–440. doi: 10.1111/j.1432-1033.1979.tb04258.x. [DOI] [PubMed] [Google Scholar]
  8. Humphries P., Old R., Coggins L. W., McShane T., Watson C., Paul J. Recombinant plasmids containing Xenopus laevis globin structural genes derived from complementary DNA. Nucleic Acids Res. 1978 Mar;5(3):905–924. doi: 10.1093/nar/5.3.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Katz L., Williams P. H., Sato S., Leavitt R. W., Helinski D. R. Purification and characterization of covalently closed replicative intermediates of ColEl DNA from Escherichia coli. Biochemistry. 1977 Apr 19;16(8):1677–1683. doi: 10.1021/bi00627a024. [DOI] [PubMed] [Google Scholar]
  10. Mansson P. E., Carter D. B., Silverberg A. B., Tully D. B., Harris S. E. Isolation and partial purification of the major abundant class rat seminal vesicle poly(A+)-messenger RNA. Nucleic Acids Res. 1979 Nov 24;7(6):1553–1565. doi: 10.1093/nar/7.6.1553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Monahan J. J., McReynolds L. A., O'Malley B. W. The ovalbumin gene. In vitro enzymatic synthesis and characterization. J Biol Chem. 1976 Dec 10;251(23):7355–7362. [PubMed] [Google Scholar]
  13. Månsson P. E., Harris S. E. Complexity of poly(A)-containing heterogeneous nuclear ribonucleic acid from mouse embryoid bodies (OTT6050). Biochemistry. 1979 May 15;18(10):2073–2078. doi: 10.1021/bi00577a036. [DOI] [PubMed] [Google Scholar]
  14. Nilson J. H., Barringer K. J., Convey E. M., Friderici K., Rottman F. M. Ontogeny of pituitary hormone mRNAs in the bovine fetus. Quantitation of pre-prolactin mRNA as a function of gestation. J Biol Chem. 1980 Jun 25;255(12):5871–5878. [PubMed] [Google Scholar]
  15. Norgard M. V., Emigholz K., Monahan J. J. Increased amplification of pBR322 plasmid deoxyribonucleic acid in Escherichia coli K-12 strains RR1 and chi1776 grown in the presence of high concentrations of nucleoside. J Bacteriol. 1979 Apr;138(1):270–272. doi: 10.1128/jb.138.1.270-272.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ostrowski M. C., Kistler M. K., Kistler W. S. Purification and cell-free synthesis of a major protein from rat seminal vesicle secretion. A potential marker for androgen action. J Biol Chem. 1979 Jan 25;254(2):383–390. [PubMed] [Google Scholar]
  17. Pan Y. C., Silverberg A. B., Harris S. E., Li S. S. Complete amino acid sequence of a major secretory protein from rat seminal vesicle. Int J Pept Protein Res. 1980 Aug;16(2):143–146. doi: 10.1111/j.1399-3011.1980.tb02947.x. [DOI] [PubMed] [Google Scholar]
  18. Parker M. G., White R., Williams J. G. Cloning and characterization of androgen-dependent mRNA from rat ventral prostate. J Biol Chem. 1980 Jul 25;255(14):6996–7001. [PubMed] [Google Scholar]
  19. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  20. Roop D. R., Tsai M. J., O'Malley B. W. Definition of the 5' and 3' ends of transcripts of the ovalbumin gene. Cell. 1980 Jan;19(1):63–68. doi: 10.1016/0092-8674(80)90388-8. [DOI] [PubMed] [Google Scholar]
  21. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  22. Stein J. P., Catterall J. F., Woo S. L., Means A. R., O'Malley B. W. Molecular cloning of ovomucoid gene sequences from partially purified ovomucoid messenger RNA. Biochemistry. 1978 Dec 26;17(26):5763–5772. doi: 10.1021/bi00619a025. [DOI] [PubMed] [Google Scholar]
  23. Tóth M. A quantitative evaluation of the effect of testosterone on RNA metabolism in the seminal vesicle of the rat. Acta Biochim Biophys Acad Sci Hung. 1978;13(1-2):23–34. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES