摘要
本文以布氏菌19株国际标准菌种为对照,对国内外实验室参考菌种14株及地方疑难菌种11株结合常规方法进行了DNA G+Cmol％测定,同时以羊种菌M16 DNA为标准,与布氏菌所有种和型的24个代表株及8株地方疑难菌种进行了DNA-DNA杂交测定,布氏菌及地方疑难菌种的G+Cmol％值范围为54.1～59.3,M16株与布氏菌参考菌种的杂交率81.7～109％,与地方疑难菌种的杂交率除一株为72.1外,其余为84.2～109％,属高度同源,鉴定为布氏菌。本试验首次使用进口新仪器“PEKIN LAMBDA 17UV/Vis SPECTROPHOTOMETER”采用热变性法测DNA G+Cmol％,复性速率法进行DNA-DNA杂交,使杂交的DNA样品变性和复性在同一比色杯中进行,省去半导体点温计操作带来的麻烦,减小了实验误差,缩短测时,结果精确、操作简便、重复性好。
Determination of bacteria DNA G+C mol% of brucella strains including 14 of laboratory reference strains and 11 strains under tests isolated in Cbina was performed as contrast with that of international reference strains, combined with routine tests. The values of G + C mol% were from 54.1 to 59.3. Applying the tests of DNA-DNA hybridization, a total of 24 strains representing all species and biovar of Brucella and 8 strains of local doubted strains were genetically very similar by the standards of DNA of M16 strains. The values of hybridization of Brucella strains and local doubted strains were 81.7~109 and 84.2~109, respectively, except for one strain 72.1. These strains were highly related and determined as Brucellas.
In this paper, the measurement of the values of DNA G + C mol% were done by using thermal denaturation and DNA-DNA hybridization were performed by using rena-turation rate method by means of a new instrument 'PEKIN-ELMER LAMBDA 17 uv/ vis SPECTRO-PHOTOMETER'. The denaturation and renaturation of DNA samples used for hybridizaton can be made in the same light path cuvette. It can avoid the inconvenience in using the thermistor thermometer and shorten the time of measuring each sample and reduce the experimental errors. It makes the results more accurate and the operation easier and thus has good reptition.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
1993年第4期334-340,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
关键词
布鲁氏菌
测定
杂交
DNA
Brucella, G + C mol% determination, DNA-DNA hybridization
