重组原核表达载体pQE30-HPV58L1的构建及鉴定

CONSTRUCTION AND IDENTIFICATION OF RECOMBINANT PLASMID PQE30-HPV58L1

目的 :构建重组原核表达载体以获得HPV58L1活性蛋白 ,为进一步研制HPV58基因工程疫苗打下基础。方法 :用聚合酶链反应 (PCR)扩增HPV58L1完整编码区基因 ,将PCR扩增产物克隆至 pUC1 9质粒中并测序。利用 pQE30质粒载体构建重组原核表达载体 pQE30 HPV58L1 ,并通过酶切电泳验证重组结果的正确性。结果 :PCR扩增出 1 6Kb特异性片段 ,经克隆至pUC1 9后测序表明序列同源性与Gen Bank报道一致。重组质粒 pQE30 HPV58L1酶切后显示其大小约 5 1Kb ,酶切图谱与预期相同。 结论 :成功构建了重组原核表达载体 pQE30 HPV58L1。

Objective:To construct a recombinant prokaryotic expression vector pQ30 HPV58L1 for obtaining the HPV58 L1 protein expressed in E coli and developing the genetic engineering vaccine.Methods:The full length L1 coden region of HPV58 was amplified by PCR,and cloned into pUC19,sequenced.The recombinant expression vector pQE30 HPV58L1 was then constructed by inserting the HPV58 L1 gene into pQE30.Restriction analysis was used to confirm the exact ligation of pQE30 HPV58L1 Results:A 1 600 bp DNA fragment was amplified with PCR.Recombinant plasmid pUC19 HPV58L1 and pQE30 HPV58L1 were constructed and sequence analysis revealed the same homology to published data in GenBank.Restriction analyses of pQE30 HPV58L1 were consistent with the theoretically predicted results.Conclusion:We successfully constructed the recombinant prokaryotic expression vector pQE30 HPV58L1