發(fā)布時(shí)間：2018-07-28 06:53

【摘要】： 目的:構(gòu)建大鼠膠質(zhì)細(xì)胞源性營養(yǎng)因子(GDNF)及內(nèi)皮素B受體(EDNRB)基因共表達(dá)腺病毒并觀察其在神經(jīng)干細(xì)胞(NSC)中的表達(dá)。方法:利用Ad-Easy腺病毒表達(dá)系統(tǒng)構(gòu)建GDNF及EDNRB基因共表達(dá)腺病毒PAd-GE,體外感染NSC后通過觀察綠色熒光蛋白表達(dá)及RT-PCR、實(shí)時(shí)熒光定量PCR.流式細(xì)胞儀法鑒定外源基因的表達(dá)。結(jié)果:NSC經(jīng)重組腺病毒PAd-GE感染后24h可觀察到綠色熒光蛋白(GFP)表達(dá),RT-PCR、實(shí)時(shí)熒光定量PCR、流式細(xì)胞儀法證實(shí)外源性GDNF及EDNRB基因均可在NSC中表達(dá)。結(jié)論:成功構(gòu)建GDNF及EDNRB基因共表達(dá)重組腺病毒,證實(shí)了其在NSC的表達(dá),為先天性巨結(jié)腸的聯(lián)合基因治療打下基礎(chǔ)。
[Abstract]:Aim: to construct rat glial cell-derived nutrition factor (GDNF) and endothelin B receptor (EDNRB) gene co-expressing adenovirus and observe its expression in neural stem cell (NSC). Methods: GDNF and EDNRB co-expressed adenovirus PAd-GE was constructed by using Ad-Easy adenovirus expression system. The expression of green fluorescent protein and RT-PCR were observed after NSC infection in vitro. The expression of exogenous gene was identified by flow cytometry. Results Twenty-four hours after infection with recombinant adenovirus PAd-GE, the expression of green fluorescent protein (GFP) was observed, and real-time fluorescence quantitative PCR was detected. Flow cytometry confirmed that exogenous GDNF and EDNRB genes could be expressed in NSC. Conclusion: the recombinant adenovirus of GDNF and EDNRB gene was successfully constructed, which confirmed its expression in NSC and laid the foundation for combined gene therapy of Hirschsprung's disease.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R656.9;R346

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 賈延R，

本文編號(hào)：2149265