Construction of Sika deer spleen cell cDNA library and discovery of four expressed sequence tags

Objective To explore and protect the gene resources of Sika deer in China in oroder to develop recombinant peptide drugs derived from Sika deer genes. Methods The total RNA was isolated from Sika deer spleen cells by using the Trizol reagent and the total mRNA was purified, complement DNA(cDNA) and the second DNA were obtained by means of reverse transcriptase PCR(RT PCR) and this cDNA was used as the template respectively. The two strains of DNA was subcloned into the pSPORT1. The recombinant plasmids of pSPORT1 were transformed into the E.coli by means of electric transcription. The cDNA library was obtained. The positive recombinant clone was selected and the sequences of EST S were analyzed. Results Sika deer spleen cell cDNA library was established successfully; the clones in it was sequenced, during the process some expresion sequence tags were discovered that shared high homology with known cDNAs deposited in the GenBank. EST 1 was homologous to human retinoblastoma binding protein 4. The protein sequence of EST 2 was 26% homologous to human vitamin K dependent protein precursor. Conclusion Some valuable gene fragments from Sika deer spleen cell cDNA library were obstained.