人乳头瘤病毒16型完整E6开放读码框的基因克隆和鉴定

GENE CLONING AND IDENTIFYING OF HUMAN PAPILLOMAVIRUS TYPE 16 COMPLETE E 6 OPEN READING FRAME

利用聚合酶链反应（ＰＣＲ）技术从人乳头瘤病毒１６型（ＨＰＶ１６）基因组中扩增得到５５５ｂｐ的Ｅ６ＤＮＡ片段，并重组到载体ｐＧＥＭＥＸＴＭ－１中，构建克隆扩增质粒ｐ１６Ｅ６－Ｇ，将此重组质粒转化大肠杆菌ＪＭ１０９，经菌落原位杂交鉴定和限制性酶切图谱分析，证实该产物为Ｅ６基因完整开放读码框（ＯＲＦ），从而为获得Ｅ６ＯＲＦ基因片段提供了一个高效。

HPV16E 6 complete ORF of a 555 bp DNA fragment has been directly amplified by PCR technic from HPV16 chromosomal DNA and cloned in the plasmid pGEMEX TM 1 to construct the sequencing and expressing plasmid p16E 6 G. Then,the plasmid p16E 6 G was transformed into E.coli JM109. Clone in situ hybridization selection and restrictive mapping analysis indicated that the 555 bp amplified PCR products were exactly the HPV16E 6 complete ORF,which provide a experimental material to express E 6 protein in eukaryotic cell expressing system.