摘要
目的克隆中国人组织激肽释放酶基因 ,为开展基因治疗高血压研究奠定基础。方法提取人胰腺组织总RNA ,逆转录后进行PCR扩增。将PCR产物回收、插入质粒KS ,酶切鉴定后双向测序分析。结果本实验克隆的激肽释放酶基因与GenBank报告的激肽释放酶基因相比 ,有一个碱基不同 ,同源性为 99.8%。结论克隆的中国人组织激肽释放酶基因可用于基因治疗等深入研究工作。
Objective To clone the human tissue kallikrein gene in Chinese. Methods Total RNA was extracted from human pancreas and human tissue kallikrein (KK) gene cDNA was amplified by PCR after reverse-transcription using Oligo(dT) primer. The original kallikrein cDNA was recovered and inserted into the Sma I site of KS plasmid. After restriction endonuclease digestion analysis, KK cDNA was sequenced by ABI 377 sequencer. Results\ The cloned kallikrein gene is about 832 bp in length. A sequence comparison of this with the reported KK sequence in the Genbank indicated that the cloned Chinese kallikrein gene has one nucleotide difference . The coded amino acid is Asp in the Genbank reported gene, while the coding amino acid of Chinese kallikrein gene is Asn. Conclusion The cloned kallikrein gene can be used for further study in gene therapy and other study.
出处
《广东医学》
CAS
CSCD
2001年第4期291-292,共2页
Guangdong Medical Journal
