鸡毒支原体抗体识别抗原的基因克隆和序列分析

抗体筛选阳性克隆，用EcoRI酶切回收3．6kb外源片段，测序。序列用大肠杆菌密码进行读框分析，至少有6个开放放阅读框，而在MG可能分为3个开放阅读框，同源性分析，发现序列的某些碱基主要与肺炎支原体、生殖支原体、山羊支原体和莱氏无胆甾原体的某些地方同源性高、没有同其他细胞或病毒有同源性的地方，说明克隆的基因完全是一个新的基因片段。

CLONING AND SEQUENCING OF THE GENE OF ANTIGEN RECOGNIZED BY ANTIBODY

The DNA of recombinant phage was digested with restriction endonuclease of EcoRI.A 3 6 kb fragment was obtained.The 3 6 kb fragment was ligated with EcoRI digested and dephosphorylated pGEM 3ZF and sequenced.There were at least 6 open reading frames in the fragment based on sequence analysis with the standard codon of E.coli.The fragment had probably 3 open reading frames in the genome of MG.Computer analysis revealed homology,to some extent,between the 3 6 kb fragment and some regions of genome in Mycoplasma pneumoniae,M.genitalium,M.capricolum and Acholeplasma laidlawii.There was no homology between the 3 6 kb fragment and genome of bacteria or virus,this shows that the 3 6 kb fragment is exactly a new gene fragment.