O'Brien M A, Roberts M S, Taghert P H
Department of Anatomy and Neurobiology, Washington University Medical School, St. Louis, Missouri 63110.
Genetics. 1994 May;137(1):121-37. doi: 10.1093/genetics/137.1.121.
We have analyzed the FMRFamide neuropeptide gene region of Drosophila melanogaster. This gene maps to the 46C region of chromosome 2R; this interval previously was not well characterized. For this genetic and molecular analysis, we have used X-ray mutagenesis, EMS mutagenesis, and the recently reported local P element transposition method. We identified four overlapping deletions, two of which have proximal breakpoints that define a 50-60-kb region surrounding the FMRFamide gene in 46C. To this small region, we mapped three lethal complementation groups; 10 additional lethal complementation groups were mapped to more distal regions of 46CD. One of these groups corresponds to even-skipped, the other 12 are previously unidentified. Using various lines of evidence we excluded the possibility that FMRFamide corresponds to any of the three lethal complementation groups mapping to its immediate 50-60-kb vicinity. The positions of two of the three lethal complementation groups were identified with P elements using a local transposition scheme. The third lethal complementation group was excluded as being FMRFamide mutants by sequence analysis and by immunocytochemistry with proFMRFamide precursor-specific antibodies. This analysis has (1) provided a genetic map of the 46CD chromosomal region and a detailed molecular map of a portion of the 46C region and (2) provided additional evidence of the utility of local transposition for targeting nearby genes.
我们分析了黑腹果蝇的FMRF酰胺神经肽基因区域。该基因定位于2R染色体的46C区域;此前该区间的特征并不明确。为了进行这项遗传和分子分析,我们使用了X射线诱变、EMS诱变以及最近报道的局部P因子转座方法。我们鉴定出四个重叠缺失,其中两个具有近端断点,这些断点定义了46C中围绕FMRF酰胺基因的一个50 - 60 kb区域。在这个小区域,我们定位了三个致死互补群;另外10个致死互补群定位于46CD的更远端区域。其中一个群对应于偶数缺失,另外12个是此前未鉴定的。通过各种证据,我们排除了FMRF酰胺对应于定位于其紧邻的50 - 60 kb区域内的三个致死互补群中任何一个的可能性。利用局部转座方案,通过P因子确定了三个致死互补群中两个的位置。通过序列分析以及使用FMRF酰胺前体特异性抗体进行免疫细胞化学,排除了第三个致死互补群是FMRF酰胺突变体的可能性。这项分析(1)提供了46CD染色体区域的遗传图谱以及46C区域一部分的详细分子图谱,(2)提供了局部转座用于靶向附近基因的实用性的更多证据。