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目的:体外扩增、鉴定并克隆间日疟原虫环子孢子蛋白(CSP)基因约772hP的基因片段,包容CSP基因的Ⅰ区、中央9肽重复区、重复后可变区及Ⅱ区.方法:采用PCR法扩增出CSP目的基因片段,以低熔点琼脂糖回收纯化,并以限制性内切酶BstNI酶切鉴定后,采用T载体连接方案,插入中间载体PUC19的多克隆位点,构建重组体PUC19/CSP,并转化大肠杆菌JM107,蓝白菌斑初筛阳性重组子,并以PCR法与限制性酶切分析对阳性克隆进一步鉴定.结果:从间日疟患者血样核酸提取物中扩增出约772hPDNA条带,而正常人血与空白对照无特异性扩增条带,经酶切鉴定证实为CSP基因片段;所构建的PUC/CSP重组体阳性克隆经双酶切与PCR鉴定与预期结果一致.结论:体外成功扩增并克隆间日疟原虫CSP基因片段,为下一步的基因序列分析及CS诊断抗原的制备打下基础.“,”AIM Amplifying and cloning the CSP gene fragment of Plasmodium vivax,it spanned the conserved region I,the central immunodominant repeat region,the variable region behind the repeats and the conserved region Ⅱ. METHODS The CSP gene fragment was amplified by PCR. After purification and idendification by di gestion with restriction enzyeme BstN I,the gene fragment was ligated with plasmid PUC19 at polylinker. The re combinant plasmid PUC19/CSP was transformed into E. colt JM107. Positive clones were screened and identified by PCR techniche and digestion with restriction enzyme. RESULTS AND CONCLUSION The CSP gene frag ment of Plasmodium vivax was successfully amplified and cloned. That would make it convenient for sequence analysis and CSP diagnostic antigen expression.