人乳头瘤病毒58型L1基因在毕赤酵母中的表达及鉴定
发布时间：2018-12-31 12:26
【摘要】： 人乳头瘤病毒(Human Papil1omavirus, HPV)是一种引起人类皮肤、粘膜的乳头状瘤或疣,并可导致恶性病变的病原体。研究表明,90%宫颈癌的发生与人乳头瘤病毒感染有关。 世界各地高危型HPV的各型感染率区别较大,除HPV16是公认发病率最高之外,HPV18及HPV58也较常见,特别是HPV58在亚洲地区表现出不同寻常的流行状况。亚洲地区HPV58的感染率明显高于世界其它地区。而在我国HPV58型也是仅次于HPV16型的高危型别。目前,世界范围内对HPV58型的进一步研究较少。基于HPV58在我国子宫颈癌中的特殊位置,我们开展了HPV58型的研究工作。 本次实验我们首先以从病理组织中提取的HPV58的基因组为模板,利用PCR的方法获得主要衣壳蛋白L1,并构建到分泌型酵母表达载体pPICZαB形成整合质粒,电转GS115酵母细胞,筛选阳性重组子,选取表达量较高的菌株进行5L发酵罐的表达,表达产物经纯化后,以本实验所制备的抗体,进行Western-Blot鉴定。结果显示,在发酵液上清中有特异蛋白条带,分子量56kD左右,证明我们得到了目的蛋白。
[Abstract]:Human papillomavirus (Human Papil1omavirus, HPV) is a pathogen that causes papilloma or warts in human skin, mucous membrane and malignant lesions. Studies have shown that 90% of cervical cancer is associated with human papillomavirus infection. The infection rates of high risk type HPV in different parts of the world are quite different. In addition to HPV16, HPV18 and HPV58 are more common, especially HPV58 shows unusual prevalence in Asia. HPV58 infection rates in Asia are significantly higher than in the rest of the world. In our country, HPV58 type is second only to HPV16 type high-risk type. At present, there is little further research on HPV58 type worldwide. Based on the special position of HPV58 in cervical cancer in China, we have carried out the research of HPV58 type. In this experiment, the genome of HPV58 extracted from pathological tissue was used as template, the main capsid protein L1 was obtained by the method of PCR, and then constructed into secretory yeast expression vector pPICZ 伪 B to form an integrated plasmid, which was electroporated into GS115 yeast cells. The positive recombinant was screened and the high expression strain was selected to express in 5L fermenter. The expressed product was purified and identified by Western-Blot with the antibody prepared in this experiment. The results showed that there were specific protein bands in the supernatant of fermentation broth, and the molecular weight of the protein was about 56kD, which proved that we got the target protein.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R346;Q78;R737.33
本文编号：2396578
[Abstract]:Human papillomavirus (Human Papil1omavirus, HPV) is a pathogen that causes papilloma or warts in human skin, mucous membrane and malignant lesions. Studies have shown that 90% of cervical cancer is associated with human papillomavirus infection. The infection rates of high risk type HPV in different parts of the world are quite different. In addition to HPV16, HPV18 and HPV58 are more common, especially HPV58 shows unusual prevalence in Asia. HPV58 infection rates in Asia are significantly higher than in the rest of the world. In our country, HPV58 type is second only to HPV16 type high-risk type. At present, there is little further research on HPV58 type worldwide. Based on the special position of HPV58 in cervical cancer in China, we have carried out the research of HPV58 type. In this experiment, the genome of HPV58 extracted from pathological tissue was used as template, the main capsid protein L1 was obtained by the method of PCR, and then constructed into secretory yeast expression vector pPICZ 伪 B to form an integrated plasmid, which was electroporated into GS115 yeast cells. The positive recombinant was screened and the high expression strain was selected to express in 5L fermenter. The expressed product was purified and identified by Western-Blot with the antibody prepared in this experiment. The results showed that there were specific protein bands in the supernatant of fermentation broth, and the molecular weight of the protein was about 56kD, which proved that we got the target protein.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R346;Q78;R737.33
【参考文献】
相关期刊论文 前2条
1 仓尧卿,朱若英;人乳头瘤病毒及其疫苗的研究[J];微生物学免疫学进展;2000年04期
2 杨海宁,于修平,卞继峰,赵蔚明,贾继辉,周亚滨,栾怡,齐眉,于谨;人乳头瘤病毒58型E6蛋白对p53的作用研究[J];中华微生物学和免疫学杂志;2003年04期
，本文编号：2396578
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