摘要
目的构建RON基因跨膜区段(RONm)的反义核酸真核表达载体。方法用RTPCR技术,从人肺癌细胞提取的总RNA克隆RONmcDNA序列。然后酶切鉴定与测序分析得到RONm的T载体,再将RONm反向插入pcDNA3.1+中,酶切鉴定得到RONm反义真核表达载体。结果用RTPCR亚克隆RONm,测序结果与GenBank的RONm相应序列(X70040)一致,阅读框无改变。构建出正确的RONm反义真核表达载体。结论成功构建RONm反义核酸真核表达载体,为进一步将RON基因反义核酸作为一种肺癌基因治疗方法的研究打下了分子生物学基础。
Objective To construct the antisense nucleic acid eukaryotic expressing vector of the cDNA sequence encoding the transmembrane domain of receptor tyrosine kinase RON (RON-RTK) gene (RONm).Methods Total RNA from lung cancer cell of human lung squamous carcinoma was extracted, and the cDNA encoding RONm was cloned by reverse transcription. The sequence of the cloned cDNA was confirmed by automated DNA sequencing, PCR and restriction enzymes analysis. The cDNA fragment was ligated into BamH Ⅰ and Hind Ⅲ sites of pcDNA 3.1+vector.Results The RONm cDNA was cloned, and the sequence analysis corresponded to data from GenBank (X70040). The pcDNA 3.1 vector contained the antisense nucleic acid of RONm.Conclusion The successful construction of the antisense nucleic acid eukaryotic expressing vector of RONm might provide materials for our further research on the mechanism of the onset and gene therapy of lung cancer.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2005年第3期291-294,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
关键词
RON基因
分子克隆
反义核酸类
真核表达载体
RON gene
molecular cloning
nucleic acid, antisense
eukaryotic expressing vector
