Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1984 Apr 1;219(1):223–231. doi: 10.1042/bj2190223

Construction and characterization of a plasmid containing complementary DNA to mRNA encoding the N-terminal amino acid sequence of the rat glutathione transferase Ya subunit.

J B Taylor, R K Craig, D Beale, B Ketterer
PMCID: PMC1153468  PMID: 6547043

Abstract

Free polyribosomal poly(A)-containing RNA isolated from normal rat liver was used to prepare a complementary DNA plasmid library in the Pst1 site of the plasmid pAT153 . A plasmid pGSTr155 complementary to mRNA coding for a glutathione transferase Ya subunit was selected by differential hybridization in situ and preliminary characterization was performed by hybrid-selected mRNA translation, immunoprecipitation and polyacrylamide-gel electrophoresis of the product synthesized in vitro. The nucleotide sequence of the complementary DNA contained within pGSTr155 was determined and shown to contain a single open reading frame corresponding to the first 129 amino acids of the N-terminus of the Ya subunit and a further 63 nucleotides upstream of the initiating methionine codon.

Full text

PDF
223

Images in this article

Selected References

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

  1. Allison J., Hall L., MacIntyre I., Craig R. K. The construction and partial characterization of plasmids containing complementary DNA sequences to human calcitonin precursor polyprotein. Biochem J. 1981 Dec 1;199(3):725–731. doi: 10.1042/bj1990725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bass N. M., Kirsch R. E., Tuff S. A., Marks I., Saunders S. J. Ligandin heterogeneity : evidence that the two non-identical subunits are the monomers of two distinct proteins. Biochim Biophys Acta. 1977 May 27;492(1):163–175. doi: 10.1016/0005-2795(77)90223-9. [DOI] [PubMed] [Google Scholar]
  4. Beale D., Ketterer B., Carne T., Meyer D., Taylor J. B. Evidence that the Ya and Yc subunits of glutathione transferase B (ligandin) are the products of separate genes. Eur J Biochem. 1982 Sep 1;126(3):459–463. doi: 10.1111/j.1432-1033.1982.tb06802.x. [DOI] [PubMed] [Google Scholar]
  5. Beale D., Meyer D. J., Taylor J. B., Ketterer B. Evidence that the Yb subunits of hepatic glutathione transferases represent two different but related families of polypeptides. Eur J Biochem. 1983 Dec 1;137(1-2):125–129. doi: 10.1111/j.1432-1033.1983.tb07805.x. [DOI] [PubMed] [Google Scholar]
  6. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  8. Burckhardt J., Telford J., Birnstiel M. L. Detection of labelled RNA species by contact hybridization. Nucleic Acids Res. 1979 Jul 11;6(9):2963–2971. doi: 10.1093/nar/6.9.2963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Craig R. K., Boulton A. P., Harrison O. S., Parker D., Campbell P. N. Studies on the intracellular segregation of polyribosome-associated messenger ribonucleic acid species in the lactating guinea-pig mammary gland. Biochem J. 1979 Sep 1;181(3):737–756. doi: 10.1042/bj1810737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Craig R. K., Brown P. A., Harrison O. S., McIlreavy D., Campbell P. N. Guinea-pig milk-protein synthesis. Isolation and characterization of messenger ribonucleic acids from lactating mammary gland and identification of caseins and pre-alpha-lactalbumin as translation products in heterologous cell-free systems. Biochem J. 1976 Oct 15;160(1):57–74. doi: 10.1042/bj1600057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Craig R. K., Hall L., Parker D., Campbell P. N. The construction, identification and partial characterization of plasmids containing guinea-pig milk protein complementary DNA sequences. Biochem J. 1981 Mar 15;194(3):989–998. doi: 10.1042/bj1940989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hall L., Craig R. K., Edbrooke M. R., Campbell P. N. Comparison of the nucleotide sequence of cloned human and guinea-pig pre-alpha-lactalbumin cDNA with that of chick pre-lysozyme cDNA suggests evolution from a common ancestral gene. Nucleic Acids Res. 1982 Jun 11;10(11):3503–3515. doi: 10.1093/nar/10.11.3503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  16. Jackson R., Hunter T. Role of methionine in the initiation of haemoglobin synthesis. Nature. 1970 Aug 15;227(5259):672–676. doi: 10.1038/227672a0. [DOI] [PubMed] [Google Scholar]
  17. Kalinyak J. E., Taylor J. M. Rat glutathione S-transferase. Cloning of double-stranded cDNA and induction of its mRNA. J Biol Chem. 1982 Jan 10;257(1):523–530. [PubMed] [Google Scholar]
  18. Ketterer B., Beale D., Meyer D. The structure and multiple functions of glutathione transferases. Biochem Soc Trans. 1982 Apr;10(2):82–84. doi: 10.1042/bst0100082. [DOI] [PubMed] [Google Scholar]
  19. Ketterer B., Beale D., Taylor J. B., Meyer D. J. The genetic relationships and inducibility of soluble glutathione transferases of the rat liver. Biochem Soc Trans. 1983 Aug;11(4):466–467. doi: 10.1042/bst0110466. [DOI] [PubMed] [Google Scholar]
  20. Ketterer B., Coles B., Meyer D. J. The role of glutathione in detoxication. Environ Health Perspect. 1983 Mar;49:59–69. doi: 10.1289/ehp.834959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Land H., Grez M., Hauser H., Lindenmaier W., Schütz G. 5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency. Nucleic Acids Res. 1981 May 25;9(10):2251–2266. doi: 10.1093/nar/9.10.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mannervik B., Jensson H. Binary combinations of four protein subunits with different catalytic specificities explain the relationship between six basic glutathione S-transferases in rat liver cytosol. J Biol Chem. 1982 Sep 10;257(17):9909–9912. [PubMed] [Google Scholar]
  23. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  24. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  25. Ramsey J. C., Steele W. J. Differences in size, structure and function of free and membrane-bound polyribosomes of rat liver. Evidence for a single class of membrane-bound polyribosomes. Biochem J. 1977 Oct 15;168(1):1–8. doi: 10.1042/bj1680001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sala-Trepat J. M., Savage M. J., Bonner J. Isolation and characterization of poly(adenylic acid)-containing messenger ribonucleic acid from rat liver polysomes. Biochim Biophys Acta. 1978 Jun 22;519(1):173–193. doi: 10.1016/0005-2787(78)90071-0. [DOI] [PubMed] [Google Scholar]
  27. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  29. Tu C. P., Weiss M. J., Karakawa W. W., Reddy C. C. Cloning and sequence analysis of a cDNA plasmid for one of the rat liver glutathione S-transferase subunits. Nucleic Acids Res. 1982 Sep 25;10(18):5407–5419. doi: 10.1093/nar/10.18.5407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tu C. P., Weiss M. J., Li N. Q., Reddy C. C. Tissue-specific expression of the rat glutathione S-transferases. J Biol Chem. 1983 Apr 25;258(8):4659–4662. [PubMed] [Google Scholar]
  31. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES