恶性疟原虫cDNA克隆(CO111)的序列测定和分析  

SEQUENCING AND ANALYSIS OF A cDNA CLONE CO111 OF PLASMODIUM FALCIPARUM 

作　　者：王燕妮[1] 黄燕[2] 李明[3] 孙万邦[1] 谢毅[2] 李英杰[3] 

机构地区：[1]遵义医学院免疫学教研室,遵义5630032 [2]复旦大学遗传工程国家重点实验室,上海2004333 [3]第一军医大学热带医学研究所,广州510515

出　　处：《中国寄生虫学与寄生虫病杂志》1999年第5期285-287,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：上海青年科技启明星计划资助; 贵州省卫生厅优秀青年科技人才基金

摘　　要：目的: 测定和分析cDNA 克隆(CO111)与抗恶性疟原虫兔血清和1株单克隆抗体呈阳性反应的序列。方法:采用PCR扩增CO111 克隆的cDNA 片段,纯化后经Klenow 酶补平,与M13m p18 载体连接,转化到大肠杆菌(E.coli) JM109。PCR筛选和鉴定M13单链,PEABI373ADNA自动测序仪测序,PC/GENE和GenBank (EM-BL)等软件进行序列分析和比较。结果:CO111 cDNA克隆含有233对核苷酸对。与GenBank 数据库中恶性疟原虫DNA比较,未发现与该cDNA相同的序列。结论:分离出1AIM: To sequence and analyse a cDNA clone CO111 , reacting with immune sera obtained from rabbits immunized with Plasmodium falciparum and one McAb against P falciparum. METHODS: cDNA clone was amplified by PCR. The PCR product was purified and polished with Klenow enzyme and ligated into the M13mp18 vector (digestd by SamI) and then transformed into E coli JM109. The positive recombinant was screened out by PCR and sequenced by the PRISM Dye Primer Sequencing Kit(ABI).The sequence was analyzed by the program from Geneva University and was compared by GenBank of EMBL. RESULTS: The nucleotide sequence of this cDNA clone contains an open reading frame of 233 bp, which encodes a predicted polypeptide of 77 amino acid residues. The ratio between A+T and G+C is 3.16. The polypeptide is highly hydrophilic and flexible. Comparison among cDNA of P falciparum from GenBank of EMBL showed that no sequence identical to this cDNA was found. CONCLUSION: A novel cDNA clone reacted with the antibodies against P falciparum was isolated.

关 键 词：恶性疟原虫 CO111 抗原 亲水性 易曲性 

分 类 号：R382.31[医药卫生—医学寄生虫学]