用CPN60基因序列研究隐孢子虫种系发育关系

畜牧兽医学报 ›› 2007, Vol. 38 ›› Issue (11): 1230-1234.

用CPN60基因序列研究隐孢子虫种系发育关系

王进产; 张龙现;娄飞;宁长申;菅复春;卢庆斌

1河南农业大学牧医工程学院，郑州 450002；2郑州牧业工程高等专科学校，郑州 450011

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-11-25 发布日期:2007-11-25

Phylogenetic Analysis of the Cryptosporidia Isolates Based on CPN60 Gene Sequence

WANG Jin-chan;ZHANG Long-xian;LOU Fei;NING Chang-shen;JIAN Fu-chun; LU Qing-bin

1College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China；2College of Zhengzhou Animal Husbandry Engineering,Zhengzhou 450011,China

Received:1900-01-01 Revised:1900-01-01 Online:2007-11-25 Published:2007-11-25

摘要/Abstract

摘要： 本试验以编码线粒体功能性蛋白Chaperonin 60 （CPN60）的核基因作为研究对象，对隐孢子虫分离株CPN60基因进行扩增测序，用Clustal X1.81对扩增序列与GenBank相关参考序列进行比对，然后用PAUP4.0程序中邻接法（Neighbor-joining，NJ）、最大简约法（Parsimony, MP）构建基因树，同时用TREEPUZZLE程序Version 4.1构建最大似然树（Maximum likelihood, ML），以确定不同隐孢子虫虫株之间的进化关系，并以18S rRNA和HSP70基因构建的进化树作参照，评价CPN60是否更适合作为隐孢子虫基因分型和进化关系的分子标记。结果显示：基于CPN60构建的进化树将隐孢子虫分为两大类：C. baileyi和C. meleagridis处于一个分枝，C. hominis、C. suis、C. parvum牛基因型和C. parvum鼠基因型处于另一个分枝上。不同隐孢子虫之间的同源性介于96％～100％，能有效区分隐孢子虫不同基因型。因此，CPN60基因序列也可作为隐孢子虫分离株种系发育的遗传标记。

关键词: 隐孢子虫, CPN60, 种系发育

Abstract: The mitochondrial functional protein Chaperonin 60（CPN60）gene was chose as the target to research the phylogenetic relationship between Cryptosporidium strains.The gene of all Cryptosporidium strains that preserved in our lab were amplified and sequenced. Then the sequences were compared to other related reference sequences with Clustal X1.81.Three methods of phylogenetic analysis were used to assess genetic relationships among Cryptosporidia isolates and the related sequences downloaded from GenBank. The distance-based neighbor-joining(NJ) analysis, parsimony(MP) analysis in PAUP4.0 were used to construct NJ tree and MP tree, while ML tree was made by using PUZZLE Version4.1.The phylogenetic relationship of different Cryptosporidium species or genotypes were established and then these phylogenetic trees were contrasted with that drew from 18S rRNA and HSP70 gene sequences, to determine whether the CPN60 gene is suitable for phylogenetic analysis of Cryptosporidium. Cryptosporidium species were divided into two groups in those phylogenetic trees, one group contained C. baileyi and C. meleagridis, the other group contained C. hominis，C. suis，C. parvum cattle genotype and C. parvum mouse genotype. The sequence identity of different Cryptosporidium species ranged from 96% to 100%. The results showed that CPN60 gene is equally suitable for the phylogentic analysis of Cryptosporidium.

Key words: Cryptosporidium, CPN60, phylogenetic development

王进产;张龙现;娄飞;宁长申;菅复春;卢庆斌. 用CPN60基因序列研究隐孢子虫种系发育关系[J]. 畜牧兽医学报, 2007, 38(11): 1230-1234.

WANG Jin-chan;ZHANG Long-xian;LOU Fei;NING Chang-shen;JIAN Fu-chun;LU Qing-bin. Phylogenetic Analysis of the Cryptosporidia Isolates Based on CPN60 Gene Sequence[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2007, 38(11): 1230-1234.

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