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Isolation of a cDNA clone of the 14-kDa subunit of the signal recognition particle by cross-hybridization of differently primed polymerase chain reactions


, : Isolation of a cDNA clone of the 14-kDa subunit of the signal recognition particle by cross-hybridization of differently primed polymerase chain reactions. Proceedings of the National Academy of Sciences of the United States of America 86(24): 9747-9751

Using an enhancement of the polymerase chain reaction (PCR) technique, we have isolated a complementary DNA encoding SRP14 (14-kDa subunit), one of six proteins contained in the signal recognition particle (SRP). Several pools of degenerate oligonucleotides encoding different peptide sequences of SRP14 were used to generate amplified DNA by the PCR. A cross-hybridization procedure was developed to identify the authentic SRP14 cDNA clone among the amplified DNA products obtained by PCR. The basis of this approach is the assumption that a partial cDNA of SRP14 should be the only DNA product common to two amplification reactions primed with different degenerate oligonucleotide mixtures. The partial canine cDNA of SRP14 identified by this procedure served as a probe to isolate a complete cDNA clone of SRP14 from a mouse embryonic cDNA library in lambda phage gt10.

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Accession: 001871950

PMID: 2557625

DOI: 10.2307/34732

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Related references

Strub, K.; Walter, P., 1989: Isolation of a cDNA clone of the 14-kDa subunit of the signal recognition particle by cross-hybridization of differently primed polymerase chain reactions. Using an enhancement of the polymerase chain reaction (PCR) technique, we have isolated a complementary DNA encoding SRP14 (14-kDa subunit), one of six proteins contained in the signal recognition particle (SRP). Several pools of degenerate oligon...

Strub K.; Walter P., 1989: Isolation of a complementary dna clone of the 14 kda subunit of the signal recognition particle by cross hybridization of differently primed polymerase chain reactions. Using an enhancement of the polymerase chain reaction (PCR) technique, we have isolated a complementary DNA encoding SPR14 (14-kDa subunit), one of six proteins contained in the signal recognition particle (SRP). Several pools of degenerate oligon...

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Lindstrom, J.T.; Chu, B.; Belanger, F.C., 1993: Isolation and characterization of an Arabidopsis thaliana gene for the 54 kDa subunit of the signal recognition particle. The first step in the routing of newly synthesized proteins into the secretory pathway is the binding of the nascent signal sequence to the signal recognition particle. The mammalian signal recognition particle is a complex consisting of 6 protein...

Lindstrom, J.; Chu, B.B.langer, F., 1993: Isolation and characterization of an Arabidopsis thaliana gene for the 54 kDa subunit of the signal recognition particle. The first step in the routing of newly synthesized proteins into the secretory pathway is the binding of the nascent signal sequence to the signal recognition particle. The mammalian signal recognition particle is a complex consisting of 6 protein...

Bovia, F.; Bui, N.; Strub, K., 1994: The heterodimeric subunit SRP9/14 of the signal recognition particle functions as permuted single polypeptide chain. The targeting of nascent polypeptide chains to the endoplasmic reticulum is mediated by a cytoplasmic ribonucleoprotein, the signal recognition particle (SRP). The 9 kD (SRP9) and the 14 kD (SRP14) subunits of SRP are required to confer elongation...

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