中文摘要：

本实验主要是从人乳腺癌细胞中克隆Rab7基因并进行基因序列比对和分析。通过培养乳腺癌细胞MCF-7和MDA-MB231,提取细胞总RNA,逆转录为cDNA,以此cDNA为模板,根据GenBank公布的人Rab7全长序列设计引物,扩增Rab7基因的开放阅读框。将其与真核表达载体pc-DNA3.1连接,构建pcDNARab7重组质粒。测序后发现MCF-7细胞中克隆的Rab7序列与GenBank序列的同源性为91%,发生了多处突变,其中94.4%为同义突变,5.5%为错义突变。但是MDA-MB231细胞中克隆的Rab7基因序列与GenBank中的Rab7序列的同源性为100%。由于发现Rab7在MCF-7乳腺癌细胞中发生突变,而在MDA-MB-231乳腺癌细胞中基因未发生突变,我们推测Rab7基因的突变是否与乳腺癌的产生有关系,这个发现为以后研究Rab7与乳腺癌的关系奠定了基础。

英文摘要：

In this experiment, we clone the Rab7 gene from human breast cancer cell lines, sequence and analysis the Rab7 gene. Cancer cell line MCF-7 and MDA-MB-231 cells were cultured, and total cellular RNA was extracted and reversely transcripted to cDNA. Using the cDNA as template, the open reading frame of Rab7 gene was amplified by the primers designed according to the sequence of Rab7 published in GenBank. Using eukaryotic expression vector pc-DNA3.1 to construct the recombinant plasmid of pcDNA-Rab7. Sequence analysis showed that the Rab7 gene sequence cloned from MCF-7 is 91% homologous with the sequence of Rab7 in GenBank. Many mutations a ppear in the sequence, including 94.4% synonymous mutations, 5.5% missense mutations. Howeve, Rab7 gene sequence cloned from MDA-MB231 cells is 100% homologous to the sequence in GenBank. Rab7 gene is mutated in MCF7 cells while not in MDA-MB-231 cells. So we think whether the mutation of Rab7 gene is associate with the generation of breast cancer. This study may lay the foundation for the further study of the relationship between breast cancer and Rab7.