水牛bFGF基因的克

发布时间：2018-06-22 17:47

本文选题：生物化学及分子生物学 + 水牛　；参考：《中国科技论文》2017年18期

【摘要】：对水牛碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)基因进行克隆、序列分析,构建bFGF表达载体,为其在水牛干细胞中的功能研究奠定基础。以水牛胎儿组织为材料提取总RNA,利用逆转录PCR(reverse transcription-PCR,RT-PCR)克隆得到的bFGF基因编码区序列,进行生物信息学分析,构建表达bFGF的逆转录病毒载体(pMXs-bFGF-IRESGFP),并在293T细胞和水牛胎儿成纤维细胞(buffalo fetal fibroblast,BFF)上进行重组载体表达鉴定。结果表明,水牛bFGF基因编码区全长468bp,编码155个氨基酸,含有纤维细胞生长因子(fibroblast growth factor,FGF)家族特有的蛋白结构;逆转录病毒载体介导的bFGF基因能在293T细胞和BFF中表达。本研究克隆出水牛bFGF编码序列,构建的逆转录病毒载体能在不同真核细胞中表达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.
【作者单位】：广西大学亚热带农业生物资源保护与利用国家重点实验室;
【基金】：高等学校博士学科点专项科研基金资助项目(20134501120003) 国家自然科学基金资助项目(31360287) 广西省自然科学基金资助项目(2014GXNSFBA118074,2015GXNSFAA139080)
【分类号】：Q78;S823.83

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