用PCR技术建立CD23全基因的无性繁殖系

THE CLONING OF HUMAN CD23 GENE BY USING POLYMERASECHAIN REACTION

参照国外已发表的ＣＤ２３ｃＤＮＡ全基因序列，自行设计并合成一对ＤＮＡ引物（２５ｍｅｒ和１８ｍｅｒ），以ＣＤ２３阳性细胞株ＨＦＢｄ２／３ＤＮＡ为模板，应用ＰＣＲ技术，成功地扩增了ＣＤ２３ｃＤＮＡ全基因（９８７ｂｐ，含人工设计的酶切点），并定向克隆入原核表达载体ｐＢＶ２２０，构建了ｐＢｖ２２０－ＣＤ２３全基因重组体，经６种限制性内切酶酶切鉴定，结果与已发表序列的酶切图谱完全一致，初步证实为ＣＤ２３全基因。这将为今后ＣＤ２３基因的序列测定和表达，进一步在基因水平上探讨ＣＤ２３的生物学功能提供了物质基础。

ith reference to the published seqtience of the human CD23 gene,a pair of DNA primers(25 bases and 18 bases)were designed and synthesized.A 987 base pair fragment containing fullCD23 cDNA gene was successfully amplified by using the polymerase chain reaction(PCR),and then cloned into pBV220 vector,constructing a recombinant plnsmid-pBCD976.It wouldpromote the further research in molecular biology of CD23。