人乳頭瘤病毒58型L1基因在畢赤酵母中的表達(dá)及鑒定
發(fā)布時(shí)間：2018-12-31 12:26
【摘要】： 人乳頭瘤病毒(Human Papil1omavirus, HPV)是一種引起人類皮膚、粘膜的乳頭狀瘤或疣,并可導(dǎo)致惡性病變的病原體。研究表明,90%宮頸癌的發(fā)生與人乳頭瘤病毒感染有關(guān)。 世界各地高危型HPV的各型感染率區(qū)別較大,除HPV16是公認(rèn)發(fā)病率最高之外,HPV18及HPV58也較常見,特別是HPV58在亞洲地區(qū)表現(xiàn)出不同尋常的流行狀況。亞洲地區(qū)HPV58的感染率明顯高于世界其它地區(qū)。而在我國(guó)HPV58型也是僅次于HPV16型的高危型別。目前,世界范圍內(nèi)對(duì)HPV58型的進(jìn)一步研究較少。基于HPV58在我國(guó)子宮頸癌中的特殊位置,我們開展了HPV58型的研究工作。 本次實(shí)驗(yàn)我們首先以從病理組織中提取的HPV58的基因組為模板,利用PCR的方法獲得主要衣殼蛋白L1,并構(gòu)建到分泌型酵母表達(dá)載體pPICZαB形成整合質(zhì)粒,電轉(zhuǎn)GS115酵母細(xì)胞,篩選陽(yáng)性重組子,選取表達(dá)量較高的菌株進(jìn)行5L發(fā)酵罐的表達(dá),表達(dá)產(chǎn)物經(jīng)純化后,以本實(shí)驗(yàn)所制備的抗體,進(jìn)行Western-Blot鑒定。結(jié)果顯示,在發(fā)酵液上清中有特異蛋白條帶,分子量56kD左右,證明我們得到了目的蛋白。
[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 偽 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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R346;Q78;R737.33
本文編號(hào)：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 偽 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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R346;Q78;R737.33
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 倉(cāng)堯卿,朱若英;人乳頭瘤病毒及其疫苗的研究[J];微生物學(xué)免疫學(xué)進(jìn)展;2000年04期
2 楊海寧,于修平,卞繼峰,趙蔚明,賈繼輝,周亞濱,欒怡,齊眉,于謹(jǐn);人乳頭瘤病毒58型E6蛋白對(duì)p53的作用研究[J];中華微生物學(xué)和免疫學(xué)雜志;2003年04期
，本文編號(hào)：2396578
本文鏈接：http://sikaile.net/yixuelunwen/binglixuelunwen/2396578.html
最近更新
教材專著
