Beeman R W, Stuart J J, Haas M S, Friesen K S
U.S. Grain Marketing Research Laboratory, U.S. Department of Agriculture, Manhattan, KS 66502, USA.
J Hered. 1996 May-Jun;87(3):224-32. doi: 10.1093/oxfordjournals.jhered.a022989.
We used a balancer chromosome to recover ethylmethanesulfonate-induced recessive mutations in a targeted region of the genome of the red flour beetle (Tribolium castaneum) by the technique of chromosome extraction. The experiments reported herein constitute the first successful application of this powerful technique in the order Coleoptera. Using the balancer chromosome maxillopedia-Dachs3 (mxpDch-3), we recovered seven recessive visible variants representing seven distinct loci and several dozen recessive lethal variants representing at least five distinct loci after screening 1,607 EMS-mutagenized chromosomes. A subset of the mxpDch-3-extracted mutations were positioned on the map of the second linkage group by a series of two-, three-, and four-point crosses. The orientation of the homeotic gene complex (HOM-C) on this linkage group was also determined. With the advent of better and more varied balancer chromosomes and the concomitant improvement of chromosome extraction procedures for genetic analysis of T. castaneum, saturation mutagenesis of targeted regions of the genome is now feasible in this species.
我们使用平衡染色体，通过染色体提取技术在赤拟谷盗（Tribolium castaneum）基因组的目标区域中恢复甲磺酸乙酯诱导的隐性突变。本文报道的实验是该强大技术在鞘翅目中的首次成功应用。使用平衡染色体巨颚-达克斯3（mxpDch-3），在筛选了1607条经甲磺酸乙酯诱变的染色体后，我们获得了代表7个不同位点的7个隐性可见变异体以及代表至少5个不同位点的几十种隐性致死变异体。通过一系列两点、三点和四点杂交，将mxpDch-3提取的一部分突变定位在第二连锁群的图谱上。同时还确定了该连锁群上同源异型基因复合体（HOM-C）的方向。随着更好、更多样化的平衡染色体的出现以及随之而来的用于赤拟谷盗遗传分析的染色体提取程序的改进，现在在该物种中对基因组目标区域进行饱和诱变是可行的。
