IN-VIVO AND IN-VITRO STABILITY OF THE BROAD-HOST-RANGE CLONING VECTOR PBBR1MCS IN 6 BRUCELLA SPECIES
IN-VIVO AND IN-VITRO STABILITY OF THE BROAD-HOST-RANGE CLONING VECTOR PBBR1MCS IN 6 BRUCELLA SPECIES
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DOI：
10.1006/plas.1995.1006
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发表时间：
1995-01-01
期刊：
影响因子：
2.6
通讯作者：
ROOF, RM
中科院分区：
文献类型：
--
作者：
ELZER, PH;KOVACH, ME;ROOF, RM
先前的研究表明，广泛宿主范围的质粒pBBR1MCS可用于流产布鲁氏菌的遗传互补。为了扩展这些观察结果，在目前公认的布鲁氏菌属六种物种中评估了 pBBR1MCS 的体内和体外稳定性。 pBBR1MCS 很容易被引入通过电穿孔测试的所有菌株中，并且在 10 天内的 5 次连续传代期间稳定地维持在肉汤培养物中，无需抗生素选择。此外，感染后 1 周从 BALB/c 小鼠脾脏中获得的所有六种含有 pBBR1MCS 的布鲁氏菌菌株的分离株都保留了该质粒。尽管 pBBR1MCS 保留了亲本质粒 pBBR1CM 中存在的动员位点，但通过接合检测 pBBR1MCS 在布鲁氏菌菌株之间转移的尝试并未成功。这些结果证明了 pBBR1MCS 在布鲁氏菌属中的体外和体内稳定性。并增强了该克隆载体对这些生物体进行遗传分析的有用性。 (C) 1995 学术出版社
Previous studies have shown that the broad-host-range plasmid pBBR1MCS can be used for genetic complementation in Brucella abortus. To extend these observations, the in vivo and in vitro stability of pBBR1MCS was evaluated in the six currently recognized species of the genus Brucella. pBBR1MCS was readily introduced into all of the strains tested by electroporation and was stably maintained in broth cultures without antibiotic selection during five serial passages over a 10-day period. Furthermore, isolates of all six Brucella strains containing pBBR1MCS obtained from the spleens of BALB/c mice 1 week postinfection maintained the plasmid. Although pBBR1MCS maintains the mobilization locus present in the parental plasmid pBBR1CM, attempts to detect transfer of pBBR1MCS between Brucella strains by conjugation were unsuccessful. These results demonstrate the in vitro and in vivo stability of pBBR1MCS in Brucella spp. and reinforce the usefulness of this cloning vector for the genetic analysis of these organisms. (C) 1995 Academic Press, Inc.