国际病毒学杂志
國際病毒學雜誌
국제병독학잡지
INTERNATIONAL JOURNAL OF VIROLOGY
2012年
4期
156-160
,共5页
张锦前%池频频%任娜%李贲%吴淑玲%温少芳%王琦%刘顺爱%成军
張錦前%池頻頻%任娜%李賁%吳淑玲%溫少芳%王琦%劉順愛%成軍
장금전%지빈빈%임나%리분%오숙령%온소방%왕기%류순애%성군
乙型肝炎表面抗原大蛋白%C53%HepG2%共定位%结合区域
乙型肝炎錶麵抗原大蛋白%C53%HepG2%共定位%結閤區域
을형간염표면항원대단백%C53%HepG2%공정위%결합구역
LHBs%C53%HepG2%co-localization%interacting region
目的 研究肝癌细胞系HepG2中乙型肝炎表面抗原大蛋白(hepatitis B virus large surface protein,LHBs)与C53蛋白是否存在共定位,查明LHBs与C53蛋白的结合区域.方法 分别构建pEGFP-C1-LHBs和pDsRed1-N1-C53质粒,共同转染肝癌细胞系HepG2,激光共聚焦观察它们在细胞内的共定位.构建质粒pCDNA-3.1(-)myc/his-LHBs和pCMV-5a-C53,并将C53基因截短构建质粒pCMV-5a-C53N、pCMV-5a-C1、pCMV-5a-C2和pCMV-5a-C3,将pCDNA-3.1(-)myc/his-LHBs和pC-MV-5a-C53及各截短质粒共转染HepG2细胞,免疫共沉淀方法验证LHBs结合的C53区域.结果 成功构建pEGFP-C1-LHBs、pDsRed1-N1-C53、pCDNA-3.1(-)myc/his-LHBs、pCMV-5a-C53、pCMV-5a-C53N、pCMV-5a-C1、pCMV-5a-C2和pCMV-5a-C3质粒；用激光共聚焦的方法证实了LHBs与C53在HepG2细胞内存在共定位；免疫共沉淀方法证实了在HepG2细胞内LHBs与C53的结合区域为其C1区.结论 LHBs与C53在肝癌细胞系HepG2内存在明确的相互作用,但仍需做进一步的功能研究.
目的 研究肝癌細胞繫HepG2中乙型肝炎錶麵抗原大蛋白(hepatitis B virus large surface protein,LHBs)與C53蛋白是否存在共定位,查明LHBs與C53蛋白的結閤區域.方法 分彆構建pEGFP-C1-LHBs和pDsRed1-N1-C53質粒,共同轉染肝癌細胞繫HepG2,激光共聚焦觀察它們在細胞內的共定位.構建質粒pCDNA-3.1(-)myc/his-LHBs和pCMV-5a-C53,併將C53基因截短構建質粒pCMV-5a-C53N、pCMV-5a-C1、pCMV-5a-C2和pCMV-5a-C3,將pCDNA-3.1(-)myc/his-LHBs和pC-MV-5a-C53及各截短質粒共轉染HepG2細胞,免疫共沉澱方法驗證LHBs結閤的C53區域.結果 成功構建pEGFP-C1-LHBs、pDsRed1-N1-C53、pCDNA-3.1(-)myc/his-LHBs、pCMV-5a-C53、pCMV-5a-C53N、pCMV-5a-C1、pCMV-5a-C2和pCMV-5a-C3質粒；用激光共聚焦的方法證實瞭LHBs與C53在HepG2細胞內存在共定位；免疫共沉澱方法證實瞭在HepG2細胞內LHBs與C53的結閤區域為其C1區.結論 LHBs與C53在肝癌細胞繫HepG2內存在明確的相互作用,但仍需做進一步的功能研究.
목적 연구간암세포계HepG2중을형간염표면항원대단백(hepatitis B virus large surface protein,LHBs)여C53단백시부존재공정위,사명LHBs여C53단백적결합구역.방법 분별구건pEGFP-C1-LHBs화pDsRed1-N1-C53질립,공동전염간암세포계HepG2,격광공취초관찰타문재세포내적공정위.구건질립pCDNA-3.1(-)myc/his-LHBs화pCMV-5a-C53,병장C53기인절단구건질립pCMV-5a-C53N、pCMV-5a-C1、pCMV-5a-C2화pCMV-5a-C3,장pCDNA-3.1(-)myc/his-LHBs화pC-MV-5a-C53급각절단질립공전염HepG2세포,면역공침정방법험증LHBs결합적C53구역.결과 성공구건pEGFP-C1-LHBs、pDsRed1-N1-C53、pCDNA-3.1(-)myc/his-LHBs、pCMV-5a-C53、pCMV-5a-C53N、pCMV-5a-C1、pCMV-5a-C2화pCMV-5a-C3질립；용격광공취초적방법증실료LHBs여C53재HepG2세포내존재공정위；면역공침정방법증실료재HepG2세포내LHBs여C53적결합구역위기C1구.결론 LHBs여C53재간암세포계HepG2내존재명학적상호작용,단잉수주진일보적공능연구.
Objective To identify the co-localization and interacting region of the hepatitis B virus large surface protein (LHBs) and C53 protein in HepG2 cell line.Methods The plasmids pEGFP-C1-LH-Bs,pDsRed1-N1-C53,pCDNA-3.1 (-) myc/his-LHBs,pCMV-5a-C53,pCMV-5a-C53N,pCMV-5a-C1,pCMV-5a-C2 and pCMV-5a-C3 were constructed. The plasmids pEGFP-C1-LHBs and pDsRed1-N1-C53were co-transfected into the HepG2 cells,and their co-localization was observed by laser scanning confocal microscope(LSCM).The plasmids pCDNA-3.1 (-) myc/his-LHBs and pCMV-5a-C53,pCMV-5a-C53N,pCMV-5a-C1,pCMV-5a-C2,pCMV-5a-C3 respectively were co-transfected into the HepG2 cells,and the interacting region of LHBs and C53 was identified by co-immunoprecipitation (CO-IP).Results The plasmids pEGFP-C1-LHBs,pDsRed1-N1-C53,pCDNA-3.1(-) myc/his-LHBs,pCMV-5a-C53,pCMV-5a-C53N,pCMV-5a-C1,pCMV-5a-C2 and pCMV-5a-C3 were constructed successfully.The co-localization of LHBs and C53 in HepG2 cells were identified,and the interacting region of LHBs and C53 was C1 of C53.Conclusion The interaction between LHBs and C53 was clear,and the further function research should be done.
