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目的 获得人趋化因子RANTES基因,用于抗艾滋病转基因中药的培育。方法 RT-PCR扩增、目的基因片段与T-载体连接和有用基因的序列分析。结果 通过淋巴细胞激活、RNA分离和RT-PCR扩增,已获得预期的276bp片段,并经PCR和酶切反应进行了鉴定。初步测序结果表明,克隆基因片段包含了编码RANTES基因信号肽与成熟蛋白的完整序列以及起始和终止密码,但编码信号肽的-10位密码子(GTT)与已发表的序列(GCT)不同。这一差异是来自基因本身的突变抑或为实验误差尚有待进一步查明。结论 克隆了人RANTES基因并进行了测序。
Objective To obtain human chemokine RANTES gene for the development of anti-AIDS transgenic Chinese medicine. Methods RT-PCR amplification, target gene fragment and T-vector connection and sequence analysis of useful genes. Results The expected 276 bp fragment was obtained by lymphocyte activation, RNA isolation and RT-PCR amplification and was identified by PCR and restriction enzyme digestion. The preliminary sequencing results showed that the cloned gene fragment contains the complete sequence encoding the signal peptide and mature protein of RANTES gene as well as the start and stop codons, but the -10 codon (GTT) encoding the signal peptide differs from the published sequence (GCT) . This difference is derived from the mutation of the gene itself or for experimental error remains to be further identified. Conclusion The human RANTES gene was cloned and sequenced.