人乳铁蛋白基因的克隆及基因序列多态性分析

Cloning of human lactoferrin gene and analysis of the polymorphism in its sequence

目的克隆人乳铁蛋白基因并进行基因序列分析.方法从人乳腺组织中提取出的总RNA进行逆转录,生成cDNA,以其为模板进行PCR扩增人乳铁蛋白cDNA.PCR产物纯化后连接到pGEM-T easy载体上并转化到DH5α菌中,然后对其进行DNA序列测定.结果本实验获得的基因序列与发表在GeneBank的hLF序列比较其同源性达99%,长度相符,为2149bp.与其他地区或国家相比较,有4～9个碱基有所不同.结论乳铁蛋白存在着基因序列多态性.

Objective To clone the gene of human lactoferrin and analyze its sequence. Methods The total RNA extracted from normal human breast tissue was processed by RT - PCR to produce hLF eDNA. The product, after being purified, was connected to pGEM - T easy vector. The conjugate was transferred into B. coli cell line, and the positive clones were screened for sequence analysis. Results Comparison of the sequence of the hLF eDNA obtained with the material published in the genebank showed： there was a 99% homology, the consistant length was 2149 bp, but 4 - 9 bp were found to be different from what were reported by other workers. Conclusion hLF eDNA has url ymorphism in sequence. Its cloning will contribute to the research work on human lactoferrin.