kar交配法转移YACs至新宿主的研究

STUDY ON THE TRANSFERRING OF YACs TO NEW HOST BY MEANS OF KAR CROSS

利用改进的kar交配法,将一个含有340kb人基因组DNA的YAC片段的供体酵母菌株YAC23与受体菌株YLB504进行交配,以选择平板对所形成的候选YAC导入菌进行筛选。经PCR分析,候选YAC导入菌在404bp处有一个扩增带,即具有受体菌株的交配型(MAT α)。进一步用脉冲电泳进行核型鉴定,证实YACs己成功地进入受体,实现了YACs从一个宿主到另一个宿主之间的转移。

The donor yeast strain YAC 23 containing a 340 kb human genomic DNA fragment in YAC (Yeast Artificial Chromosome) was mated with recipient strain YLB 504 by means of improved kar cross and the condidate YACductants were assayed by PCR and the amplification band of 404 bp indicated that they were the same as those of recipient strains (i. e. MATα) Further analysis of the electro-karyotype of the candidate YACductants by PFGE (Pulsed Field Gel Electrophore-sis) confirmed that the YACs had succeeded in entering into the recipient cells. The YACs had thus completed the process of transferring from one host to the other.