Molnar C M, Reece T, Williams J A, Bell J B
Department of Genetics, University of Alberta, Edmonton, Canada.
Genome. 1988 Apr;30(2):211-7. doi: 10.1139/g88-036.
P-element mediated transformation was utilized to introduce a suppressor tRNA gene (Sup3e tRNA(UGASER)) from Schizosaccharomyces pombe into Drosophila melanogaster. Thirteen independently transformed lines were characterized as to the number of cytological locations of the transposons. It was ascertained that the suppressor tRNA gene of interest was introduced into each transformed strain. The helper P element used (p pi 25.1) allows further transposition to occur, and it was determined that from one to seven copies of the heterologous tRNA(UGASER) gene per strain were present among the respective transformed strains. The number of transposons per transformed line was established by in situ hybridization to salivary gland chromosomes as well as by Southern hybridization analyses and there was good agreement in the totals determined by these two techniques.
利用P因子介导的转化方法,将来自粟酒裂殖酵母的一个抑制性tRNA基因(Sup3e tRNA(UGASER))导入黑腹果蝇。对13个独立转化的品系进行了转座子细胞学定位数量的鉴定。确定每个转化菌株都导入了感兴趣的抑制性tRNA基因。所使用的辅助P因子(p pi 25.1)允许进一步发生转座,并且确定在各个转化菌株中,每个菌株存在1到7个拷贝的异源tRNA(UGASER)基因。通过对唾液腺染色体进行原位杂交以及Southern杂交分析,确定了每个转化品系中转座子的数量,这两种技术确定的总数吻合良好。