Molecular cloning of chromosome I DNA from @entity375 : isolation and characterization of the XXXX gene and adjacent regions of the chromosome.

@entity72194 @entity27961 @entity16791 @entity375
Molecular cloning techniques were used to isolate and characterize the DNA including and surrounding the @entity16791 and @entity27961 genes on the left arm of chromosome I of the @entity375 @entity375 . A plasmid that complemented a temperature-sensitive @entity16791 mutation was isolated from a @entity375 genomic DNA library in a shuttle vector. Plasmids containing @entity27961 -complementing DNA were obtained from other investigators. Several lines of evidence (including one-step gene replacement experiments) demonstrated that the complementing plasmids contained the bona fide @entity16791 and @entity27961 genes. These sequences were then used to isolate additional DNA from chromosome I by probing a @entity375 genomic DNA library in a lambda vector. A total of 28 kilobases (kb) of contiguous DNA surrounding the @entity16791 and @entity27961 genes was isolated, and a restriction map was determined. Electron microscopy of R-loop-containing DNA and RNA blot hybridization analyses indicated that an 18-kb segment contained at least seven transcribed regions, only three of which corresponded to previously known genes ( @entity16791 , @entity27961 , and @entity72194 ). Southern blot hybridization experiments suggested that none of the genes in this region was duplicated elsewhere in the @entity375 genome. The centers of @entity16791 and @entity27961 were only approximately 7.5 kb apart, although the genetic map distance between them is approximately 13 centimorgans. As previous studies with @entity375 have indicated that 1 centimorgan generally corresponds to approximately 3 kb, the region between @entity16791 and @entity27961 appears to undergo meiotic recombination at an unusually high frequency.
