CAJ | 학술논문

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方法扩增出一1.17 Kb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S26／3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。
장자행분리、전대배양적내몽고지구산양유산의원체안상규방법분리순화，제취의원체기인조DNA작위모판，안조국외발표적의원체주요외막단백（MOMP）기인량단서렬설계합성일대인물，용PCR방법확증출일1.17 Kb적DNA편단。이용인물상예선설계적한제성내절매위점，장확증편단경한제성내절매절할후련접도pUC19질립상응위점상，전화대장간균DH5α，사선중조자。경PCR검측화내절매분석감정함MOMP기인적중조자질립。대극륭편단진행전서렬분석，결과증명득도MOMP전편마서렬적기인극륭。본주의원체MOMP편마구유1170개핵감산조성。서렬비교발현본주의원체적MOMP기인여국외적양유산의원체S26／3주적MOMP기인완전상동，여B577주적MOMP기인부유일개핵감산적동의변이。
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．