中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2009年
1期
71-76
,共6页
潘晖榕%李少伟%刘波%朱子恒%曹欢欢%顾颖%程通%王颖彬%张军%夏宁邵
潘暉榕%李少偉%劉波%硃子恆%曹歡歡%顧穎%程通%王穎彬%張軍%夏寧邵
반휘용%리소위%류파%주자항%조환환%고영%정통%왕영빈%장군%하저소
人乳头状瘤病毒6型%大肠杆菌表达系统%纯化%类病毒颗粒%免疫原性
人乳頭狀瘤病毒6型%大腸桿菌錶達繫統%純化%類病毒顆粒%免疫原性
인유두상류병독6형%대장간균표체계통%순화%류병독과립%면역원성
Human papillomavirus type 6%Escherichia coli expression system%Purification%Virus-like particle%Immunogenicity
目的 利用大肠杆菌表达系统制备人乳头状瘤病毒6型(human papillomavirus type 6,HPV-6)类病毒颗粒(virus-like particles,VLP)并研究其免疫原性.方法 在大肠杆菌ER2566中表达HPV-6 L1蛋白,并以硫酸铵沉淀、离子交换色谱、疏水相互作用色谱等手段对其进行纯化.纯化后的HPV-6 L1经体外组装形成VLP后以动态光散射,透射电镜对其形态进行检测,并以假病毒中和实验评价HPV-6 L1 VLP在实验动物体内所诱导的抗HPV-6/11中和抗体水平.结果 HPV-6 L1蛋白在大肠杆菌中以可溶形式表达,经过纯化后的HPV-6 L1蛋白可以在体外组装为半径25 nm左右的VLP.该VLP可以在山羊及兔体内诱导高滴度的HPV-6/11中和抗体.结论 大肠杆菌表达系统可以简便高效制备具有免疫原性的HPV-6 VLP,可以用于HPV-6疫苗的研究.
目的 利用大腸桿菌錶達繫統製備人乳頭狀瘤病毒6型(human papillomavirus type 6,HPV-6)類病毒顆粒(virus-like particles,VLP)併研究其免疫原性.方法 在大腸桿菌ER2566中錶達HPV-6 L1蛋白,併以硫痠銨沉澱、離子交換色譜、疏水相互作用色譜等手段對其進行純化.純化後的HPV-6 L1經體外組裝形成VLP後以動態光散射,透射電鏡對其形態進行檢測,併以假病毒中和實驗評價HPV-6 L1 VLP在實驗動物體內所誘導的抗HPV-6/11中和抗體水平.結果 HPV-6 L1蛋白在大腸桿菌中以可溶形式錶達,經過純化後的HPV-6 L1蛋白可以在體外組裝為半徑25 nm左右的VLP.該VLP可以在山羊及兔體內誘導高滴度的HPV-6/11中和抗體.結論 大腸桿菌錶達繫統可以簡便高效製備具有免疫原性的HPV-6 VLP,可以用于HPV-6疫苗的研究.
목적 이용대장간균표체계통제비인유두상류병독6형(human papillomavirus type 6,HPV-6)류병독과립(virus-like particles,VLP)병연구기면역원성.방법 재대장간균ER2566중표체HPV-6 L1단백,병이류산안침정、리자교환색보、소수상호작용색보등수단대기진행순화.순화후적HPV-6 L1경체외조장형성VLP후이동태광산사,투사전경대기형태진행검측,병이가병독중화실험평개HPV-6 L1 VLP재실험동물체내소유도적항HPV-6/11중화항체수평.결과 HPV-6 L1단백재대장간균중이가용형식표체,경과순화후적HPV-6 L1단백가이재체외조장위반경25 nm좌우적VLP.해VLP가이재산양급토체내유도고적도적HPV-6/11중화항체.결론 대장간균표체계통가이간편고효제비구유면역원성적HPV-6 VLP,가이용우HPV-6역묘적연구.
Objective To produce human papillomavirus type 6(HPV-6)virus-like particles with Escherichia coli expression system and study its immunogenicity.Methods HPV-6 L1 gene was inserted into pmkaryotic expression vector pTO-T7 and then expressed in Escherichia coli ER2566.The HPV-6 L1 protein was purified by ammonium sulfate precipitation,ion-exchange chromatography,and hydrophobic interaction chromatography.Then the purified HPV-6 L1 self-assembled into virus-like particle after removing 1,4dithiothreitol(DTr).The morphology of the virus-like particles was investigated with dynamic light scatter and transmission electron microscopy,and the immunogenicity was determined with in vitro pseudownons neutralization as8ay.Results HPV-6 L1 was expressed in soluble form in Escherichia coli.Following the removal of DTT,purified HPV-6 L1 protein could assemble into virus-like particles as 25 am in the radius.And the animal immunization test showed HPV-6 virus-like particles can elite hish titer neutralizing antibodies.Conclusion The bacterially expressed HPV-6 L1 VLP is highly immunogenieity and easy to produce.And it can be good candidate of HPV-6 vaccine.
