Method for cloning genes

Details Method for cloning genes Method for cloning genes

1980-12-18

1983-07-19

Tanenholtz, Alvin E.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 68, 435 70, 435172, 435 91, 436 63, C12Q 168, C12P 2100, C12P 2102, C12N 1500, C12P 1934, G01N 3350

Patent

active

043944434

ABSTRACT:
A method is provided for isolating and identifying a recombinant clone having a DNA segment therein coding for at least one desired heterologous polypeptide, at least a short amino acid sequence of which is known, by effecting cDNA synthesis on a mixture of mRNAs containing the mRNA coding for the desired polypeptide, isolating the resultant cDNA mixture, inserting the resultant cDNA into recombinant cloning vehicles, transforming hosts with the vehicles, separating the transformants and isolating and identifying a recombinant clone containing a DNA segment which is homologous over at least a portion thereof to at least one oligonucleotide probe specific for the DNA segment; wherein the probe is an extension of the nucleotide sequence of an oligonucleotide primer having a nucleotide sequence complementary to a region of the target mRNA coding for a portion of the known amino acid sequence, and is complementary to a longer region of the target mRNA coding for a longer portion of the known amino acid sequence. Recombinant clones coding for human histocompatibility antigens are provided, in particular, clones for HLA-B antigens.

REFERENCES:
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Houghton et al.; Nucl. Acids. Res. 8, 1913 (1980).
Montgomery et al., Cell 14, 673 (1978).
Wallace et al.; Nucl. Acids. Res. 6, 3543 (1979).
Kennell; Progr. Nucl. Acid Res. Mol. Biol. 11, 259 (1971).
Orr et al.; Proc. Natl. Acad. Sci. USA 76, 4395 (1979).
Riggs et al., Am. J. Hum. Genet. 31, 531-538 (1979).
Hidde L. Ploegh, et al., Proc. Natl. Acad. Sci. USA, Oct. 1980, vol. 77, No. 10, pp. 6081-6085, Molecular Cloning of a Human Histocompatibility Antigen cDNA Fragment.
F. Sanger, et al., Proc. Natl. Acad. Sci. USA, Dec. 1977, vol. 73, No. 12, pp. 5463-5467, DNA Sequencing with Chain-Terminating Inhibitors.
David Zimmeran, et al., Proc. Natl. Acad. Sci USA, Sep. 1978, vol. 75, No. 9, pp. 4257-4261, 3'-Terminal Nucleotide Sequence of Encephalomyocarditis Virus RNA Determined by Reverse Transcriptase and Chain-Terminating Inhibitors.
Jack W. Szostak et al., Nature, Jan. 1977, vol. 265, pp. 61-63, Specific Binding of a Synthetic Oligodeoxyribonucleotide to Yeast Cytochrome c mRNA.
Barbara E. Noyes, et al., Proc. Natl. Acad. Sci. USA, Apr. 1979, vol. 76, No. 4, pp. 1770-1774, Detection and Partial Sequence Analysis of Gastrin mRNA by Using an Oligodeoxynucleotide Probe.

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