布氏菌544A基因片段的PCR扩增及其序列分析

CLONING AND SEQUENCING OF BRUCELLA 544A GENE FRAGMENT

利用ＰＣＲ和ＤＮＡ重组技术，成功的扩增了牛布氏菌（Ｂｒ．ａｂｏｒｔｕｓ）５４４Ａ基因。回收含Ｂｒ．ａｂｏｒｔｕｓ５４４ＡＤＮＡ片段，插入ＳｍａＩ单酶切的ｐＵＣ９载体中，进行核苷酸序列分析，证实克隆的ＤＮＡ片段长度为２２４ｂｐ，在该序列中含有单个开放阅读框架。将重组子用Ｋｐｎ１和ＢａｍＨ１双酶切下Ｂｒ．ａｂｏｒｔｕｓ５４４ＡＤＮＡ片段标记探针，均与６种布氏菌杂交出现阳性结果。说明Ｂｒ．ａｂｏｒｔｕｓ５４４Ａ２２４ｂｐＤＮＡ片段是６个菌种间的高度保守序列，具有很高的同源性。为布氏病临床医学和流行病学调查研究奠定了基础。

A DNA fragment of Brucella abortus 544A has been successfully amplified using the technique of PCR.The gene fragment is put into the pUC 9 vector is sequensed.We prove that the length of DNA cloning fragment is 224 base pair and there is a mono opening reading frame in the fragment.A probe from Br.abortus 544A is used to hybridize with six strains of brucella,and the result are postive.It come to the conclusion that fragment of 544A is the high conservative sequence in the six Brucella strains,and there is very great homogenity each other.The study made the basis for further research of Brucella disease.