摘要
采用PCR方法,从HPV16型野毒株质粒中分离出HPV16L1(55597151bp)、HPV16E7C(682855bp)基因片段,采用PCR产物的TA克隆法,将L1、E7C分别插入pGEMTEasy载体,构建克隆载体pTAL1、pTAE7C。BamHⅠ、HindⅢ酶切pTAE7C,Bg1Ⅱ、ClaⅠ酶切pTAL1,回收L1、E7C片段,利用BamHⅠ、Bg1Ⅱ酶切产生相同粘末端的特点,产生L1E7C重组基因片段,将其克隆入质粒pBlueScriptsk,构建了pBSL1E7C重组克隆质粒,HindⅢ、XhoⅠ酶切pBSL1E7C,释放L1E7C基因片段,定向重组入pCA14腺病毒载体,成功构建真核表达载体HPV16L1E7C重组腺病毒载体:pCA14L1E7C。
The entire HPV16L1 gene and C terminal HPV16E7 gene were amplified from wild type HPV16 plasmid using PCR method.After using T A cloning of PCR products,E7C gene and L1 gene are inserted into the pGEM T easy vector.After digested pTAL1 with restriction endonuclease Bgl Ⅱ,Hind Ⅲ and digested pTAE7C with restriction endonuclease BamH Ⅰ,Cla Ⅰ,E7C and L1 gene are inserted into the polycloning site of clone vector pBlueScriptks in which E7C and L1 gene are fused .Then using Hind Ⅲ and Xho Ⅰ in polycloning site of recombinant vector pBlueScriptsk ,L1 E7C gene is released and inserted into adenovirus vector pCA14,a recombinant adenovirus expressing vector is generated.
出处
《山东医科大学学报》
1999年第1期1-5,共5页
Acta Academiae Medicinae Shandong
基金
国家自然科学基金