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本文以人乳头状瘤病毒(HPV)16型-PBR_(322)重组质粒,对大肠杆菌HB101进行转化实验。经筛选、扩增、酶切、电泳鉴定。获得满意的电泳图谱及kb值,HPV-16Bam HI DNA为7.2kb,其260/280O.D比值为1.9-2。纯度合乎要求,可用以制备探针。以HPV-16DNA7.2kb片段为探针,应用~(32)P(α)-dATP标记进行了点杂交及Southern印迹杂交实验。结果表明38例宫颈癌组织DNA,其点杂交阳性率为76.3％,Southern杂交阳性率为65.7％。点杂交与Southern杂交结果基本符合。上述结果说明了在人宫颈癌组织中,存在着HPV-16相关的基因组,Southern杂交表明其以整合形式存在。说明宫颈癌的发病可能与HPV-16型的感染有密切关系。从分子水平进一步探讨了宫颈癌的病毒病因。
In this paper, the human papillomavirus (HPV) 16 -PBR_ (322) recombinant plasmid, Escherichia coli HB101 transformation experiments. After screening, amplification, digestion, electrophoresis identification. Satisfactory electropherograms and kb values were obtained. The HPV-16Bam HI DNA was 7.2 kb with a ratio of 260/280 OD of 1.9-2. Purity is desirable and can be used to prepare probes. The HPV-16DNA7.2kb fragment was used as a probe to probe the hybridization and Southern blotting with ~ (32) P (α) -dATP. The results showed that 38 cases of cervical cancer DNA, the spot hybridization positive rate was 76.3%, Southern blot was 65.7% positive. Point hybridization and Southern hybridization results basically in line. The above results indicate that HPV-16-related genomes exist in human cervical cancer tissues, and Southern hybridization indicates that they exist in an integrated form. The incidence of cervical cancer may be closely related to HPV-16 infection. From the molecular level to further explore the etiology of cervical cancer virus.