猪附红细胞体特异性基因的克隆和序列分析

Cloning and Sequencing Analysis of the Gene Eperythrozoon Suis of Pig

为分析吉林省流行的猪附红细胞体基因序列,根据发表的猪附红细胞体16S rRNA基因序列设计引物,扩增出长度约为541 bp的基因片段,并成功地将该基因克隆到pGEM-T Easy载体。将经NotⅠ酶切鉴定和PCR鉴定为阳性的重组质粒进行测序,与发表的附红细胞体基因序列进行比较,试验所获得的核苷酸序列与猪附红细胞体16S rRNA基因序列非常相近,同源性为97.8%-100%,从而证明吉林省所流行的猪附红细胞体属于16S rRNA基因群。

To analyze the nucleotide sequence of Epertythrozoon suis of pig in Jilin province, the published sequence of the E. suis 16S rRNA gene was used for designing primers, A 541-basepair fragment was amplified, The purified PCR product was cloned into pGEM-T Easy vector. The recombinant plasmid was identified by Not Ⅰ restriction enzyme and PCR. The DNA fragment was sequenced and the sequences of nucleotide was compared with published E. Suis 16S rRNA gene sequence. The result showed that the nucleotide sequence had closely relation to E. suis 16S rRNA. The nucleic acid sequence homology was 97.8% - 100%.