MOLECULAR CLONING OF A NEW VARIANT OF HUMAN PAPILLOMAVIRUS TYPE 6 ISOLATED FROM A CHINESE WOMAN AND EXPRESSION OF ITS L1 GENE IN E. coli——MOLECULAR CLONING & EXPRESSION OF HPV6 GENE

A human papillomavirus genome DNA of 7.9 kb from a Chinese woman with genital condyloma acuminata was cloned in Bam HI site of pAT153. According to the results obtained from Southern blotting, restriction mapping as well as partial DNA sequencing, the isolated genome (HPV6BV) had obvious variance and was referred to as a new variant of HPV6 found in China the first time. HPV6BV L1 gene was successfully expressed in E. coli as a fusion protein with pUR288. The β-galactosidase/L1 fusion protein reacted with both β-galactosidase antiserum and HPV antibody using Western blot technique. The E. coli-produced fusion protein, possessing HPV antigenicity, may provide a reagent for clinical diagnosis and epidemiological survey.     

我国人乳头瘤病毒6型新变种基因组克隆及L1晚期结构基因在大肠杆菌中表达

本文从一例中国妇女外阴尖锐湿疣组织标本中提取DNA,BamHI酶切,以pAT153为载体,成功地克隆了我国人乳头瘤病毒6型(HPV-6BV)基因组.经South-ern杂交,酶谱及部分DNA序列分析,发现该克隆的HPV6基因组有明显变异,可视为我国发现的第一个HPV6的新变种.本文进一步分离了HPV6BV的L1基因,以pUR288为载体,在大肠杆菌中表达了β-半乳糖苷酶/HPV6BVL1N融合蛋白.经免疫转印试验证明,它既能与β-半乳糖苷酶抗血清反应,又能与HPV抗体反应,具有融合前各自的抗体亲和活性.该蛋白可用作诊断试剂及流行病学研究.