中国人视黄醇结合蛋白cDNA的克隆与序列分析

CLONING AND SEQUENCING OF cDNA CODING FOR RETINOL BINDING PROTEIN OF CHINESE

目的： 为了进行中国人视黄醇结合蛋白（ｈｕｍ ａｎ ｒｅｔｉｎｏｌ－ｂｉｎｄｉｎｇ ｐｒｏ－ｔｅｉｎ， ｈＲＢＰ）的原核表达，克隆出ｈＲＢＰ的ｃＤＮＡ。 方法： 利用反转录聚合酶链式反应（ＲＴ－ＰＣＲ）方法获得ｈＲＢＰ的ｃＤＮＡ，克隆入ｐＧＥＭ－Ｔ载体后，在美国ＡＢＩＤＮＡ 自动测序仪上以双脱氧法进行核苷酸序列测定。结果： 扩增出了约０．５８ｋｂ的特异片段，并筛选出阳性重组克隆质粒ｐＧＥＭ－ＲＢＰ，核苷酸序列测定表明其含有一段５８０ 个碱基的插入片段，与文献报道的一致。结论： 我们已获得了中国人ＲＢＰ的ｃＤＮＡ，可用于进行原核表达。

The study is to clone the cDNA coding for the human retinol binding protein (hRBP). Methods: Isolate and clone the cDNA coding for hRBP into a E. coli cloning vector pGEM T using RT PCR and then sequence it on American automated capillary electrophoresis genetic analyzer. Results: We amplified a 0.58kb PCR product and the positive recombinant plasmid pGEM hRBP was screened. DNA sequence analysis confirmed that the sequence of our PCR product was consisitent with the reported sequence. Conclusion: We have cloned the cDNA coding for hRBP and it is ready for our further studies.