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PMID: 388427 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Construction and selection of recombinant plasmids containing full-length complementary DNAs corresponding to rat insulins I and II.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 76 ·No. 10 ·1979-10-00 ·Pages 5036-40

Chan SJ, Noyes BE, Agarwal KL, Steiner DF

Abstract

We have used a synthetic deoxydecanucleotide to generate an insulin-specific cDNA probe suitable for selecting transformants that contain nearly full-length cDNAs corresponding to the mRNAs coding for rat insulins I and II. Double-stranded cDNA was synthesized from x-ray-induced rat insulinoma poly(A)-RNA, inserted in pBR322 plasmid DNA by the homopolymeric tailing technique, and cloned in Escherichia coli chi 1776. Colony hybridization with oligonucleotide-primed cDNA yielded 16 positive clones of which 7 corresponded to rat insulin I mRNA and 9 to rat insulin II mRNA. Restriction endonuclease maps of representative clones of each group indicated that these contained the complete coding sequences, as was confirmed by nucleotide sequence analysis of the 5' region of the cloned DNA for rat insulin II. Nucleotide sequence analysis also established the amino acid sequence of the prepeptide of rat preproinsulin II. Comparison of the amino acid sequence of the prepeptides of rat preproinsulin I and II shows that three conservative amino acid substitutions have occurred in this region of the molecule.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cloning, Molecular DNA, Recombinant/metabolism Escherichia coli/metabolism Insulin/biosynthesis Molecular Weight Nucleic Acid Hybridization Plasmids Rats
Chemicals
DNA, Recombinant Insulin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Chan S J
Noyes B E
Agarwal K L
Steiner D F
References (26)
26 references, click to expand
  1. Recombinant plasmids containing Xenopus laevis globin structural genes derived from complementary DNA.
    Nucleic Acids Res. 1978 Mar;5(3):905-24 PMID: 347404
  2. A general method for cloning eukaryotic structural gene sequences.
    Proc Natl Acad Sci U S A. 1976 Sep;73(9):3146-50 PMID: 61588
  3. Detection and partial sequence analysis of gastrin mRNA by using an oligodeoxynucleotide probe.
    Proc Natl Acad Sci U S A. 1979 Apr;76(4):1770-4 PMID: 88048
  4. Amplification and characterization of a beta-globin gene synthesized in vitro.
    Cell. 1976 Jun;8(2):163-82 PMID: 61066
  5. A transplantable insulinoma in the rat.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):628-32 PMID: 191819
  6. Partial purification and characterization of the mRNA for rat preproinsulin.
    Proc Natl Acad Sci U S A. 1976 Oct;73(10):3539-43 PMID: 185618
  7. 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):5350-4 PMID: 414220
  8. A bacterial clone synthesizing proinsulin.
    Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727-31 PMID: 358198
  9. Rat insulin genes: construction of plasmids containing the coding sequences.
    Science. 1977 Jun 17;196(4296):1313-9 PMID: 325648
  10. A new method for sequencing DNA.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4 PMID: 265521
  11. Detection of gastrin-specific mRNA using oligodeoxynucleotide probes of defined sequence.
    J Biol Chem. 1979 Aug 25;254(16):7472-5 PMID: 468766
  12. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels.
    J Mol Biol. 1977 Feb 15;110(1):119-46 PMID: 845942
  13. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose.
    Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408-12 PMID: 4504350
  14. Ribonucleic acid isolated by cesium chloride centrifugation.
    Biochemistry. 1974 Jun 4;13(12):2633-7 PMID: 4831907
  15. Enzymatic in vitro synthesis of globin genes.
    Cell. 1976 Feb;7(2):279-88 PMID: 60178
  16. Chemical synthesis of polynucleotides.
    Angew Chem Int Ed Engl. 1972 Jun;11(6):451-9 PMID: 4626260
  17. Biochemical method for mapping mutational alterations in DNA with S1 nuclease: the location of deletions and temperature-sensitive mutations in simian virus 40.
    Proc Natl Acad Sci U S A. 1975 Mar;72(3):989-93 PMID: 165498
  18. Insulin biosynthesis in the rat: demonstration of two proinsulins.
    Proc Natl Acad Sci U S A. 1969 Jan;62(1):278-85 PMID: 4890253
  19. The amino acid sequence of the insulin from a primitive vertebrate, the atlantic hagfish (Myxine glutinosa).
    J Biol Chem. 1975 Jul 10;250(13):5183-91 PMID: 1097441
  20. 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):3961-5 PMID: 1105573
  21. Cell-free synthesis of rat preproinsulins: characterization and partial amino acid sequence determination.
    Proc Natl Acad Sci U S A. 1976 Jun;73(6):1964-8 PMID: 778852
  22. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase.
    Nucleic Acids Res. 1976 Jan;3(1):101-16 PMID: 765970
  23. Replication of small plasmids in extracts of Escherichia coli.
    Mol Gen Genet. 1976 Jun 15;145(3):273-80 PMID: 781516
  24. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis.
    Biochemistry. 1975 Aug 26;14(17):3787-94 PMID: 1174504
  25. Species variation in the amino acid sequence of insulin.
    Am J Med. 1966 May;40(5):662-6 PMID: 5949593
  26. DNA sequence analysis: a general, simple and rapid method for sequencing large oligodeoxyribonucleotide fragments by mapping.
    Nucleic Acids Res. 1974 Mar;1(3):331-53 PMID: 10793670
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.76.10.5036
Published
1979-10-00
Pages
5036-40
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC413074
Subset
IM
Databases
GENBANK
J00748
External Links
PubMed source DOI: 10.1073/pnas.76.10.5036 PMC Full Text View journal details
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