Haenlin M, Steller H, Pirrotta V, Mohier E
Cell. 1985 Apr;40(4):827-37. doi: 10.1016/0092-8674(85)90342-3.
The K10 female sterility locus involved in establishment of the embryonic dorsoventral axis maps genetically to the 2E2-2F1 interval of the Drosophila X chromosome. We microdissected the 2E2-2F3 region from salivary gland chromosomes and used clones obtained from the microdissected fragments to establish a chromosomal walk covering more than 200 kb. To identify the K10 gene we used P-mediated transformation with cosmid clones constructed in cos-P, a cosmid vector incorporating the terminal repeats of the P element. Clone cos9, containing a 43 kb insert, transformed the germ line of homozygous K10 females and allowed production of normal progeny. It also rescued three genes, crooked neck, pecanex, and kurz, which map genetically near K10. Transformation experiments using smaller fragments of cos9 localize the K10+ function within 11 kb. Northern blots hybridized with probes from this region indicate the presence of several mRNA species. Each transcript has been assigned to a complementation group.
参与胚胎背腹轴建立的K10雌性不育位点在遗传学上定位于果蝇X染色体的2E2 - 2F1区间。我们从唾液腺染色体上显微切割了2E2 - 2F3区域，并使用从显微切割片段中获得的克隆构建了一个覆盖超过200 kb的染色体步移。为了鉴定K10基因，我们使用了在cos - P中构建的黏粒克隆进行P介导的转化，cos - P是一种包含P因子末端重复序列的黏粒载体。含有43 kb插入片段的克隆cos9转化了纯合K10雌性的种系，并使其产生正常后代。它还拯救了在遗传学上位于K10附近的三个基因，即弯颈、pecanex和kurz。使用cos9较小片段的转化实验将K10 +功能定位在11 kb范围内。用该区域的探针进行的Northern杂交表明存在几种mRNA种类。每个转录本都已被归入一个互补群。
