人乳铁蛋白(hLF)cDNA的克隆及序列分析

Cloning and Sequence Analysis of Human Lactoferrin

从正常人外周血中分离中性粒白细胞(WBC),提取总RNA,用RT-PCR的方法扩增人乳铁蛋白(hLF)cDNA.cDNA分hLF 1.5h、LF 0.8两段扩增并连入pMD18-T载体上,再利用限制酶连入巴氏毕赤酵母表达载体pPICZ-αA中,构成完整的hLF基因.序列测定表明,所克隆的hLF基因序列全长为2,136bp,与Gene Bank中登录的序列相比,同源性达99%以上.

White blood cells were isolated from normal human peripheral bloods and total RNA was extracted. Two fragments of human lactoferrin （hLF） were amplified by RT-PCR and then cloned into pMD18-T vector. Full length of hLF was ligated from these two fragments by restriction enzymes into pPICZα-A expression vector, hLF gene consists of 2,136bp. Sequence analysis found 99% homology with other sequences in Gene Bank.