球形芽孢杆菌Ts-1原生质体电诱导质粒转化研究

THE STUDY OF BACILLUS SPHAERICUS T(s-1) PROTOPLAST-PLASMID TRANSFORMATION OF ELECTROPORATION

本文探讨了球形芽孢杆菌Bacillus sphaericus Ts-1原生质体电诱导质粒pHV 33转化的最佳条件:3个连续的间隔为1秒,时程为10μs,强度为21KV/cm的脉冲,使其转化频率为2.44×102转化子/μg DNA,转化效率为3.16×10-6,质粒转化吸附的饱和浓度为5μg/106CFU。利用此条件,将重组杀蚊毒蛋白克隆pJB417转入Bacillus subtilis 168M和Bacillus sphaericus Ts-1中,使得B.subtilis有了杀蚊活性,但未能提高Ts-1的毒性。

This report gave the best conditions of Bacillus sphaericus Ts-1 protoplast-plasmid pHV33 clectroporation. The highest transformation frequency and transformation efficiency induced by three pulse of 21 KV/cm and 101μs duration applied at an interval of one sec., was 2.44×10~2 transformants/μg DNA and 3.16×10^(-6) respectively. The saturated concentration of DNA absorbed by the protoplast was 5μg DNA/10~9 cells/ml. By means of this method, pJB417, a recombinant mosquito larvicide clone, was introduced into B. subtilis 168M and B. sphaericus Ts-1. The transformants of B. subtilts 168M with biocide activity were obtained, but the toxicity of B. sphaericus Ts-1 was not increased.