论文部分内容阅读
将恶性疟原虫保护性抗原复合基因（PfCMR）与表达质粒pWR450—1分别经BamHI、EcoRI双酶切后回收、纯化和重组并转化大肠杆菌JM109，再经含氨苄青霉素LB培基初筛后，挑白色菌落扩增，用PCR复筛和双酶切鉴定，阳性克隆子pWR450—1—B14用IPTG诱导，在JM109中表达。表达产物经SDS—PAGE分析，在65kDa处出现较宽的蛋白条带。用SOS显色法证实此表达蛋白具有β-半乳糖苷酶活性。表达的融合蛋白与抗恶性疟原虫多克隆免疫血清特异结合．其滴度高达1：3200；Westernblot分析显示，表达蛋白能与特异性抗体产生较强的免疫反应。上述结果提示表达的融合蛋白具有生物学和免疫学活性。
Plasmodium falciparum protective antigen complex gene (PfCMR) and expression plasmid pWR450-1 were digested with BamHI and EcoRI respectively, then purified, recombinantly transformed and transformed into E. coli JM109. After screening with ampicillin LB medium, Pick white colonies amplified by PCR screening and double enzyme digestion identification, positive clones pWR450-1-B14 induced by IPTG, expressed in JM109. The expression product was analyzed by SDS-PAGE, showing a broad protein band at 65 kDa. SOS chromogenic method confirmed that the expressed protein has β-galactosidase activity. The expressed fusion protein specifically bound to the polyclonal immune serum against Plasmodium falciparum. The titer was up to 1: 3200. Western blot analysis showed that the expressed protein reacted strongly with specific antibodies. The above results suggest that the expressed fusion protein has biological and immunological activity.