恶性疟原虫组氨酸富集蛋白II基因克隆及测序

中国寄生虫学与寄生虫病杂志 ›› 1999, Vol. 17 ›› Issue (5): 7-281.

恶性疟原虫组氨酸富集蛋白II基因克隆及测序

方建民;余新炳;罗树红;徐劲

中山医科大学寄生虫学教研室广州 510089

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1999-10-30 发布日期:1999-10-30

CLONING AND SEQUENCING OF THE GENES CODING FOR THE HISTIDINE RICH PROTEIN II OF PLASMODIUM FALCIPARUM *

FANG Jianmin;YU Xinbing;LUO Shuhong;XU Jin

Department of Parasitology;Sun Yat sen University of Medical Sciences;Guangzhou 510089

Received:1900-01-01 Revised:1900-01-01 Online:1999-10-30 Published:1999-10-30

摘要/Abstract

摘要： 　　目的:比较恶性疟原虫不同分离株组氨酸富集蛋白II( HRPII)基因全编码区核苷酸序列。方法: PCR扩增恶性疟原虫海南株(FCC1/HN)和越南株(VN)的HRPII基因全编码区, 其产物经HindIII和Bam HI双酶切,定向克隆入pUC19, 采用Sanger 双脱氧链末端终止法测序。结果:FCC1/HN 株和VN 株HRPII基因均为1 020bp, 无内含子, 两者仅10 个碱基不同。FCC1/HN 株HRPII与VN 株、IMTM22 株和Itg2 株间氨基酸同源性分别为98.8% 、92.2% 和98.7% ; 4 株虫体均有拷贝数不等的丙氨酸-组氨酸-组氨酸(AHH)和丙氨酸-组氨酸-组氨酸-丙氨酸-丙氨酸-天冬氨酸(AHHAAD) 重复序列, 具有相同的信号肽和糖基化位点。4 个分离株HRPII抗原表位相同,位于易曲性较高的5端非AHH 和AHHAAD 重复区。结论: FCC1/HN 株与VN 株的HRPII高度同源,与IMTM22 株和Itg2 株存在序列差异

关键词: 恶性疟原虫, 组氨酸富集蛋白II, 序列分析

Abstract: 　AIM: To compare and analyze the homology of genes encoding histidine rich proteinII (HRPII) of different Plasmodium falciparum isolates. METHODS: Using PCR technique, the complete genes coding for HRPII of P.falciparum isolates FCC1/HN and VN isolates were amplified. PCR products were digested by HindIII/BamHI and cloned into plasmid pUC19. The recombinant plasmid HRPII/pUC19 was screened and identified by PCR and restriction analysis. The cloned HRPII genes were sequenced by Sangers method. RESULTS: HRPII genes of FCC1/HN and VN isolates were successfully amplified and cloned into pUC19. DNA sequencing showed that the coding length of HRPII gene was 1 020 bp without introns in FCC1/HN and VN isolates, however, there were ten points mutations between them. FCC1/HN isolate exhibited 98 8%, 92 2% and 98 7% homology in amino acids with isolates VN, IMTM22, and Itg2, respectively. Though the numbers of repeat sequences were different in four isolates, they had the same hydrophobic leader sequence and a single putative glycosylation site. The secondary structure analysis showed that the main antigenic determinants of four isolates were located on 5end non repeat region (amino acids 1-60). CONCLUSION: FCC1/HN isolate was highly homologous in the coding region of HRPII with VN, IMTM22, and Itg2 isolate. Four isolates exhibited similar structural characteristics and antigenic determinants in HRPII.

Key words: Plasmodium falciparum, histidine rich protein II, sequence analysis

方建民;余新炳;罗树红;徐劲. 恶性疟原虫组氨酸富集蛋白II基因克隆及测序[J]. 中国寄生虫学与寄生虫病杂志, 1999, 17(5): 7-281.

FANGJianmin;YUXinbing;LUOShuhong;XUJin . CLONING AND SEQUENCING OF THE GENES CODING FOR THE HISTIDINE RICH PROTEIN II OF PLASMODIUM FALCIPARUM *[J]. Chinese Journal of Parasitology and Parasitic Diseases, 1999, 17(5): 7-281.

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