[Isolation of new Zinc finger genes through cDNA library screening combined with RACE].
Zhanwen Du, Liren Liu, Jun‐Wu Zhang
Abstract
Zhanwen Du, Liren Liu, Jun‐Wu Zhang
Abstract
Most of the important functionally proteins contain the corresponding function domains that consist of conserved amino acid sequences. The study provided a method to identify novel genes that encode proteins containing important functionally domains with conserved sequences. First, primers were designed according to the sequence of the cDNA library vector and the ESTs that have been obtained by reverse PCR and degenerate primers encoding Zinc finger domain. The cDNA library DNA was used as template for PCR amplification. The amplified fragment that contains nonhomologous sequences of the cDNA was inserted into pGEM-T easy vector. The fragment was recovered and used as a probe for screening the cDNA library. Several cDNAs with full length that encode proteins with Zinc finger domain and represent the original ESTs have been successfully cloned from a human bone marrow cDNA library. This strategy can also be used in screening genes that encode proteins containing differential function domains with conserved sequences.
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Most of the important functionally proteins contain the corresponding function domains that consist of conserved amino acid sequences. The study provided a method to identify novel genes that encode proteins containing important functionally domains with conserved sequences. First, primers were designed according to the sequence of the cDNA library vector and the ESTs that have been obtained by reverse PCR and degenerate primers encoding Zinc finger domain. The cDNA library DNA was used as template for PCR amplification. The amplified fragment that contains nonhomologous sequences of the cDNA was inserted into pGEM-T easy vector. The fragment was recovered and used as a probe for screening the cDNA library. Several cDNAs with full length that encode proteins with Zinc finger domain and represent the original ESTs have been successfully cloned from a human bone marrow cDNA library. This strategy can also be used in screening genes that encode proteins containing differential function domains with conserved sequences.
Key concepts: Complementary DNA, cDNA library, Biology, Rapid amplification of cDNA ends, ENCODE, Zinc finger, Gene, Genomic library