摘要
采用６５ＫＤａ热激蛋白基因为模板，选择位于３９８－８３６基因序列处的一对分枝杆菌通用引物，扩增出４３９ｂｐ的基因片段。将１２种标准分枝杆菌的ＰＣＲ扩增产物即４３９ｂｐ用ＢｓｔＥⅡ限制性内切酶作酶切分析，根据酶切带型的大小、强弱以及带型数量进行分枝杆菌菌种鉴定。通过对１２种分枝杆菌的ＰＣＲ反应和限制性酶切分析，可将被检标本中大部分的分枝杆菌鉴定到种。本试验无需进行杂交或使用放射性物质，２４ｈ即可完成。
The gene encoding for the 65KDa protein was used as template to amplify a 439bp between position 398 and 836 of the published gene sequence through PCR technique. PCR products obtained with primers common to all mycobacteria were carried out restriction enzyme analysis by using one restriction enzyme, BstE . Evaluating the size,amount and intensity of the fragments does differentiation of mycobacteria to the species level. The majority species of 12 were differentiated to species by PCRrestriction enzyme pattern analysis. The procedure does not involve hybridization steps or the use of radioactivity and can be completed within 1 working day.
出处
《西北农业学报》
CAS
CSCD
1999年第2期6-10,共5页
Acta Agriculturae Boreali-occidentalis Sinica
关键词
分枝杆菌
PCR
限制性酶切分析
Mycobacteria
PCR
Restriction enzyme pattern analysis.
