PMID: 6986172Jan 1, 1980Paper

Recombinant plasmids containing avian vitellogenin structural gene sequences derived from complementary DNA

Biochimica Et Biophysica Acta
T OhnoJ P Jost

Abstract

Purified mRNA coding for chicken vitellogenin, a precursor of egg yolk proteins, was transcribed to complementary DNA (cDNAvit) with avian myeloblastosis virus (AMV) reverse transcriptase. Double-stranded cDNA was synthesized with Escherichia coli DNA polymerase I (fragment A) using the self priming ability of the cDNA. Following S1 nuclease digestion the double-stranded cDNA was inserted into the Hind III site of plasmid pBR322 using the poly(dA) . poly(dT) tailing method, and the hybrid molecules were used to transform Escherichia coli chi 1776. Ampicillin-resistant colonies were screened by colony hybridization with 125I-labeled vitellogenen mRNA. Further screening of positive clones was done by agarose gel electrophoresis and in situ hybridization with 125I-labeled vitellogenin mRNA. In addition, plasmid DNA covalently bound to diazotized paper was used to select complementary mRNA sequences. The cloned vitellogenin sequences were shown to hybridize to a mRNA which directs the synthesis of immunoprecipitable vitellogenin when translated in a reticulocyte lysate cell-free system. The length of the inserted cDNA was determined by agarose gel electrophoresis and heteroduplex mapping. The largest insertion was about 2500 base p...Continue Reading

References

Feb 1, 1979·Journal of Virology·G E HoutsJ W Beard
Sep 1, 1976·Cell·J R Tata
Aug 1, 1976·European Journal of Biochemistry·H R Pelham, R J Jackson
Feb 1, 1965·Journal of Molecular Biology·F W STUDIER

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Citations

Jul 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·A F WilksJ P Jost
Oct 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·A M SinhaM Rabinowitz
Jul 24, 1981·Nucleic Acids Research·A C ArnbergA B Geert
Mar 1, 1982·CRC Critical Reviews in Biochemistry·D Shapiro

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