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| The CDS Cloning of HCRTR1 Gene with Over-lap PCR from Qinchuan Cattle and its Expression in E.coli |
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Abstract With the Overlap-PCR methods, the 1 278 bp cds sequences of HCRTR1 gene in Qinchuan cattle was got. The fragments were cloned into the pMD18-T vector and the positive clone plasmids was sequenced. Results showed that the cloned fragments had 98% homology with the sequence of GenBank after being blasted. Digested the plasmid with BamHⅠand HindⅢ and the 1 278 bp was obtained. The obtained fragment was ligated with the longer fragment digested with the same restriction enzyme from the expression vector, pET32 a(+). The positive recombinated plasmid was transformed Bl21(DE3) and induced with IPTG. But the HCRTR1 gene cannot be expressed by E.coli. And the tranmembrane structure may explain why.
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Received: 19 September 2007
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