Kapoun A M, Kaufman T C
Howard Hughes Medical Institute, Department of Biology, Indiana University, Bloomington 47405, USA.
Genetics. 1995 Jun;140(2):643-58. doi: 10.1093/genetics/140.2.643.
We have identified regulatory regions of the homeotic gene proboscipedia that are capable of repressing a linked white minigene in a manner that is sensitive to chromosomal pairing. Normally, the eye color of transformants containing white in a P-element vector is affected by the number of copies of the transgene; homozygous flies have darker eyes than heterozygotes. However, we found that flies homozygous for select pb DNA-containing transgenes had lighter eyes than heterozygotes. Several pb DNA fragments are capable of causing this pairing sensitive (PS) negative regulation of white. Two fragments in the upstream DNA of pb, 0.58 and 0.98 kb, are PS; additionally, two PS sites are located in the second intron, including a 0.5-kb region and 49-bp sequence. This phenotype is not observed when two PS sites are located at different chromosomal insertion sites (in trans-heterozygous transgenic animals), indicating that the pb-DNA-mediated repression of white is dependent on the pairing or proximity of the PS regions. The observed phenomenon is similar to transvection in which certain alleles of a gene can complement each other, but only when homologous chromosomes are paired. Interestingly, the intronic PS regions contain positive regulatory sequences for pb, whereas the upstream PS sites contain pb negative regulatory elements.
我们已经鉴定出同源异型基因触须基因(proboscipedia)的调控区域,该区域能够以对染色体配对敏感的方式抑制与之相连的白色微型基因。正常情况下,在P因子载体中含有白色基因的转化体的眼色受转基因拷贝数的影响;纯合子果蝇的眼睛比杂合子的颜色更深。然而,我们发现,对于含有特定触须基因(pb)DNA的转基因纯合子果蝇,其眼睛比杂合子的更浅。几个触须基因(pb)DNA片段能够导致这种对白色基因的配对敏感(PS)负调控。触须基因(pb)上游DNA中的两个片段,分别为0.58 kb和0.98 kb,具有配对敏感(PS)特性;此外,两个配对敏感(PS)位点位于第二个内含子中,包括一个0.5 kb的区域和一段49 bp的序列。当两个配对敏感(PS)位点位于不同的染色体插入位点时(在反式杂合转基因动物中),未观察到这种表型,这表明触须基因(pb)DNA介导的对白色基因的抑制依赖于配对敏感(PS)区域的配对或接近程度。观察到的现象类似于异位显性,即基因的某些等位基因可以相互互补,但只有当同源染色体配对时才会发生。有趣的是,内含子配对敏感(PS)区域包含触须基因(pb)的正调控序列,而上游配对敏感(PS)位点包含触须基因(pb)的负调控元件。