本文選題：生物化學(xué)及分子生物學(xué) + 水牛��； 參考：《中國科技論文》2017年18期

【摘要】：對水牛堿性成纖維細(xì)胞生長因子(basic fibroblast growth factor,bFGF)基因進(jìn)行克隆、序列分析,構(gòu)建bFGF表達(dá)載體,為其在水牛干細(xì)胞中的功能研究奠定基礎(chǔ)。以水牛胎兒組織為材料提取總RNA,利用逆轉(zhuǎn)錄PCR(reverse transcription-PCR,RT-PCR)克隆得到的bFGF基因編碼區(qū)序列,進(jìn)行生物信息學(xué)分析,構(gòu)建表達(dá)bFGF的逆轉(zhuǎn)錄病毒載體(pMXs-bFGF-IRESGFP),并在293T細(xì)胞和水牛胎兒成纖維細(xì)胞(buffalo fetal fibroblast,BFF)上進(jìn)行重組載體表達(dá)鑒定。結(jié)果表明,水牛bFGF基因編碼區(qū)全長468bp,編碼155個(gè)氨基酸,含有纖維細(xì)胞生長因子(fibroblast growth factor,FGF)家族特有的蛋白結(jié)構(gòu);逆轉(zhuǎn)錄病毒載體介導(dǎo)的bFGF基因能在293T細(xì)胞和BFF中表達(dá)。本研究克隆出水牛bFGF編碼序列,構(gòu)建的逆轉(zhuǎn)錄病毒載體能在不同真核細(xì)胞中表達(dá)bFGF。
[Abstract]:The basic fibroblast growth factor (basic fibroblast growth) gene of buffalo was cloned and sequenced. The expression vector of bFGF was constructed, which laid a foundation for the study of its function in buffalo stem cells. Total RNAs were extracted from buffalo fetal tissues. The sequence of bFGF gene coding region was cloned by reverse transcription-PCR (RT-PCR) and analyzed by bioinformatics. A retrovirus vector expressing bFGF (pMXs-bFGF-IRESGFP) was constructed and expressed in 293T cells and buffalo fetal fibroblasts (buffalo fetal fibroblast BFF). The results showed that the encoding region of buffalo bFGF gene was 468 BP, encoding 155 amino acids and containing the unique protein structure of fibroblast growth factor (fibroblast growth) family, and the expression of bFGF gene mediated by retrovirus vector was found in 293T cells and BFF. In this study, the buffalo bFGF coding sequence was cloned, and the constructed retroviral vector was able to express bFGF in different eukaryotic cells.
【作者單位】： 廣西大學(xué)亞熱帶農(nóng)業(yè)生物資源保護(hù)與利用國家重點(diǎn)實(shí)驗(yàn)室;
【基金】：高等學(xué)校博士學(xué)科點(diǎn)專項(xiàng)科研基金資助項(xiàng)目(20134501120003) 國家自然科學(xué)基金資助項(xiàng)目(31360287) 廣西省自然科學(xué)基金資助項(xiàng)目(2014GXNSFBA118074,2015GXNSFAA139080)
【分類號】：Q78;S823.83

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