羊流产衣原体主要外膜蛋白基因的克隆与序列分析  

CLONING AND SEQUENCE ANALYSIS OF THE MAJOR OUTER MEMBRANE PROTEIN GENE OF AN OVINE ABORTION STRAIN OF CHLAMYDIA PSITTACI

作　　者：苗振川 哈斯阿古拉[2] 赵亚芳[1] 张宝发[1] 张鹤龄[2] 

机构地区：[1]内蒙古畜牧科学院,呼和浩特010030 [2]内蒙古大学生物工程中心,呼和浩特010021

出　　处：《畜牧兽医学报》2001年第1期86-91,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：内蒙古自治区自然科学基金资助

摘　　要：将自行分离、传代培养的内蒙古地区山羊流产衣原体按常规方法分离纯化 ,提取衣原体基因组DNA作为模板 ,按照国外发表的衣原体主要外膜蛋白 (MOMP)基因两端序列设计合成一对引物 ,用PCR方法扩增出一 1 17Kb的DNA片段。利用引物上预先设计的限制性内切酶位点 ,将扩增片段经限制性内切酶切割后连接到 pUC19质粒相应位点上 ,转化大肠杆菌DH5α ,筛选重组子。经PCR检测和内切酶分析鉴定含MOMP基因的重组子质粒。对克隆片段进行全序列分析 ,结果证明得到MOMP全编码序列的基因克隆。本株衣原体MOMP编码区由 1170个核苷酸组成。序列比较发现本株衣原体的MOMP基因与国外的羊流产衣原体S2 6/3株的MOMP基因完全相同 ,与B577株的MOMP基因仅有一个核苷酸的同义变异。In order to clone the major outer membrane protein(MOMP) gene of a goat abortion strain of Chlamydia psittaci isolated from Inner Mongolia area,a pair of primers were synthesized according to the published sequences of Chlamydial MOMP gene,and the genomic DNA was extracted from the purified Chlamydial particles and employed as templat,then polymerase chain reaction was performed.As a result,a 1.17 Kb DNA fragment was amplified.The PCR product was recovered from low melting point agarose gel and underwent cleavage by restriction enzymes whose recognition sites had been designed precedingly on the ends of the primers,then ligated to the corresponding site of plasmid pUC19.The ligation reaction mixture was used to trans form E.coli DH5α.The recombinant was screened by PCR detection or restriction enzymes analysis.The DNA sequencing was performed.It showed that this recombinant fragment covered the whole coding region of the MOMP gene whice was 1170 bp long.It also revealed that this MOMP gene of Chlamydial strain was just the same as that of England ovine abortion strain S26/3,and there was only one nucleotide synonymous mutation in comparison with that of strain B577.

关 键 词：羊 流产 衣原体 外膜蛋白基因 序列分析 鹦鹉热衣原体 

分 类 号：S858.235[农业科学—临床兽医学] S852.67[农业科学—兽医学]