Amplification,Clone And Sequencing Of Gene Fragment Of The Rat Oviduct Specific Glycoprotein

Objective To amplify,clone and sequence the gene fragment of rat oviduct specific glycoprotein. Methods By finding the nucleotide sequence of the oviduct specific glycoprotein of house mouse, gold mouse, pig and human, the homology was analyzed among them by Primer Premier 5.0 soft. The resylt of the homologous analysis was that the homology of the nucleotide sequence of the oviduct specific glycoprotein of house mouse and that of gold mouse,pig and human was 80%, 54% and 52% respectively. A pair of primer had been designed in the highly homologous region of them with Primer Premier 5.0 soft. The total RNA of the oviduct of the rat was extracted with TRIzol Reagent. The gene fragment had been obtained by RT-PCR with the designed primer. Linking the gene fragment to the pMD 18-T Vector, the recombinant had been obtained. The recombinant was transformed to the JM109 competent cell. The transformed JM109 competent cell was cultured and amplified in LB culture medium. The recombinant was extracted from the coliform . The gene fragment was sequenced and the homology was analyzed between the obtained sequence and the nucleotide sequence of the oviduct specific glycoprotein of house mouse, gold mouse, pig and human by Primer Premier 5.0 soft. Results A piece of gene fragment was obtained by RT-PCR with the designed primer. The sequence of the gene fragment was highly homologous with the nucleotide sequence of the oviduct specific glycoprotein of house mouse , gold mouse, pig and human. Conclusion The obtained gene sequence in this experiment is the gene fragment of the nucleotide sequence of the rat oviduct specific glycoprotein.