醫(yī)學(xué)論文范文:假型逆轉(zhuǎn)錄病毒載體高效介導(dǎo)兔平滑肌細(xì)胞基因轉(zhuǎn)染

醫(yī)學(xué)論文范文:假型逆轉(zhuǎn)錄病毒載體高效介導(dǎo)兔平滑肌細(xì)胞基因轉(zhuǎn)染

【摘要】 目的 構(gòu)建假型逆轉(zhuǎn)錄病毒載體MuLV/VSV�睪，并用于轉(zhuǎn)染兔平滑肌細(xì)胞，為兔平滑肌細(xì)胞基因轉(zhuǎn)染尋找一種高效的載體。方法 構(gòu)建含有報道基因lacZ的假型逆轉(zhuǎn)錄病毒載體MuLV/VSV�睪，測定滴度，并轉(zhuǎn)染兔平滑肌細(xì)胞，觀察其轉(zhuǎn)導(dǎo)效率。并與MuLV的轉(zhuǎn)導(dǎo)效率進(jìn)行比較。結(jié)果 構(gòu)建的MuLV/VSV�睪載體，病毒滴度為6～7.8×106CFU，轉(zhuǎn)染兔平滑肌細(xì)胞的效率是(92±12)%。而MuLV的轉(zhuǎn)導(dǎo)效率為(24±5)%。結(jié)論 成功構(gòu)建了假型逆轉(zhuǎn)錄病毒載體MuLV/VSV�睪載體，該載體作為一種高效的載體可用于兔平滑肌細(xì)胞基因轉(zhuǎn)染。

【關(guān)鍵詞】 假型逆轉(zhuǎn)錄病毒載體;平滑肌細(xì)胞;轉(zhuǎn)染

High�瞖fficiency gene transfer into rabbit smooth muscle cellsby pseudotyped retroviral vector

PEI Fei, HE Rui, LI Jun�瞴an, YU Hong

Department of Cardio�瞭horacic Surgery, the Second Affiliated Hospital,Medical School of Xi�餫n Jiaotong University, Xi�餫n 710004, China

ABSTRACT: Objective To construct pseudotyped retroviral vector MuLV/VSV�睪 and transfer it into rabbit smooth muscle cells (SMC) in order to provide a high�瞖fficiency vector for SMC gene transfer. Methods We constructed pseudotyped retroviral vector MuLV/VSV�睪 containing the previously reported gene lacZ, determined the titer, and determined the efficiency of gene transfer into SMC mediated by pseudotyped retroviral vector MuLV/VSV�睪. Finally the transfer efficiency was compared with that by MuLV. Results MuLV/VSV�睪 vector was constructed. The titer of the vector was 6-7.8×106CFU, the transfer efficiency was (92±12)% by using MuLV/VSV�睪 vector and (24±5)% by MuLV vector. Conclusion Pseudotyped retroviral vector MuLV/VSV�睪 which was constructed successfully is a kind of high�瞖fficiency gene transfer vector in smooth muscle cells醫(yī).學(xué)全.在.線網(wǎng)站gydjdsj.org.cn.

KEY WORDS: pseudotyped retroviral vector; smooth muscle cell; gene transfer

基因治療是指將具有防治潛能的外源基因，通過相應(yīng)的載體轉(zhuǎn)移到患者的有關(guān)組織器官和細(xì)胞內(nèi)，獲得適當(dāng)?shù)谋磉_(dá)達(dá)到防治和減輕疾病的目的。其中目的基因載體的選擇是基因治療的關(guān)鍵之一。逆轉(zhuǎn)錄病毒是較常用的一種病毒載體。鼠白血病病毒MuLV衍生的逆轉(zhuǎn)錄病毒載體被廣泛應(yīng)用于將基因轉(zhuǎn)染到哺乳細(xì)胞中并長期表達(dá)[1]。但該載體穩(wěn)定性差，產(chǎn)生的病毒滴度低，轉(zhuǎn)染到人類細(xì)胞效率低，并且只能轉(zhuǎn)染分裂期細(xì)胞。VSV�睪，皰疹性口炎蛋白VSV的包膜蛋白，是一種單鏈蛋白，可以完全替代MuLV的env蛋白，產(chǎn)生基于MuLV的高轉(zhuǎn)染率的病毒載體MuLV/VSV�睪，已經(jīng)被用來構(gòu)建假型MuLV/VSV�睪以提高M(jìn)uLV載體的穩(wěn)定性及轉(zhuǎn)染效率[2]。本研究用兔平滑肌細(xì)胞作為靶細(xì)胞，觀察MuLV/VSV�睪的轉(zhuǎn)導(dǎo)效率，并與MuLV的轉(zhuǎn)導(dǎo)效率進(jìn)行比較，探討MuLV/VSV�睪用于平滑肌細(xì)胞基因轉(zhuǎn)染的可行性。

1 材料與方法

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