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医学论文范文:假型逆转录病毒载体高效介导兔平滑肌细胞基因转染
【摘要】 目的 构建假型逆转录病毒载体MuLV/VSVG，并用于转染兔平滑肌细胞，为兔平滑肌细胞基因转染寻找一种高效的载体。方法 构建含有报道基因lacZ的假型逆转录病毒载体MuLV/VSVG，测定滴度，并转染兔平滑肌细胞，观察其转导效率。并与MuLV的转导效率进行比较。结果 构建的MuLV/VSVG载体，病毒滴度为6～7.8×106CFU，转染兔平滑肌细胞的效率是(92±12)%。而MuLV的转导效率为(24±5)%。结论 成功构建了假型逆转录病毒载体MuLV/VSVG载体，该载体作为一种高效的载体可用于兔平滑肌细胞基因转染。
【关键词】 假型逆转录病毒载体;平滑肌细胞;转染
Highefficiency gene transfer into rabbit smooth muscle cellsby pseudotyped retroviral vector
PEI Fei, HE Rui, LI Junyan, YU Hong
Department of Cardiothoracic Surgery, the Second Affiliated Hospital,Medical School of Xian Jiaotong University, Xian 710004, China
ABSTRACT: Objective To construct pseudotyped retroviral vector MuLV/VSVG and transfer it into rabbit smooth muscle cells (SMC) in order to provide a highefficiency vector for SMC gene transfer. Methods We constructed pseudotyped retroviral vector MuLV/VSVG containing the previously reported gene lacZ, determined the titer, and determined the efficiency of gene transfer into SMC mediated by pseudotyped retroviral vector MuLV/VSVG. Finally the transfer efficiency was compared with that by MuLV. Results MuLV/VSVG vector was constructed. The titer of the vector was 6-7.8×106CFU, the transfer efficiency was (92±12)% by using MuLV/VSVG vector and (24±5)% by MuLV vector. Conclusion Pseudotyped retroviral vector MuLV/VSVG which was constructed successfully is a kind of highefficiency gene transfer vector in smooth muscle cells医.学全.在.线网站www.lindalemus.com.
KEY WORDS: pseudotyped retroviral vector; smooth muscle cell; gene transfer
基因治疗是指将具有防治潜能的外源基因，通过相应的载体转移到患者的有关组织器官和细胞内，获得适当的表达达到防治和减轻疾病的目的。其中目的基因载体的选择是基因治疗的关键之一。逆转录病毒是较常用的一种病毒载体。鼠白血病病毒MuLV衍生的逆转录病毒载体被广泛应用于将基因转染到哺乳细胞中并长期表达[1]。但该载体稳定性差，产生的病毒滴度低，转染到人类细胞效率低，并且只能转染分裂期细胞。VSVG，疱疹性口炎蛋白VSV的包膜蛋白，是一种单链蛋白，可以完全替代MuLV的env蛋白，产生基于MuLV的高转染率的病毒载体MuLV/VSVG，已经被用来构建假型MuLV/VSVG以提高MuLV载体的稳定性及转染效率[2]。本研究用兔平滑肌细胞作为靶细胞，观察MuLV/VSVG的转导效率，并与MuLV的转导效率进行比较，探讨MuLV/VSVG用于平滑肌细胞基因转染的可行性。
1 材料与方法
