用质粒DNA对牛链球菌原生质体进行转化

Transformation of Streptococcus bovis protoplasts by plasmid DNA.

作者信息

Mareková M, Kmet V, Javorský P

机构信息

Institute of Animal Physiology, Slovak Academy of Science, Kosice, Slovakia.

出版信息

Lett Appl Microbiol. 1996 Feb;22(2):159-61. doi: 10.1111/j.1472-765x.1996.tb01132.x.

DOI:10.1111/j.1472-765x.1996.tb01132.x

PMID:8936377

Abstract

The transformation and subsequent regeneration of ruminal strain Streptococcus bovis AO24/85 protoplasts by plasmid DNA was studied. The best stabilizer for regeneration of protoplasted cells was 5% sucrose in the regeneration medium and in the agar plates. Optimal concentration of polyethylene glycol 6000 in the transformation medium was 25% for both plasmids tested. Addition of Ca2+ and Mg2+ ions (2.5 mmol l-1) to the transformation medium increased the proportion of regenerated cells. Transformation frequencies were 3 x 10(3) transformants per microgram of pNZ12 and 2.4 x 10(2) per microgram of pJK108, respectively.

摘要

研究了瘤胃菌株牛链球菌AO24/85原生质体通过质粒DNA的转化及随后的再生。再生培养基和琼脂平板中用于原生质体细胞再生的最佳稳定剂是5%蔗糖。对于所测试的两种质粒，转化培养基中聚乙二醇6000的最佳浓度均为25%。向转化培养基中添加Ca2+和Mg2+离子（2.5 mmol l-1）可增加再生细胞的比例。转化频率分别为每微克pNZ12有3×10(3)个转化子和每微克pJK108有2.4×10(2)个转化子。