Phenotype-driven is the name for an ap-proach used to study gene functions through mutagenesis, location and cloning of the mutant gene. In this study, 150 male C57BL/6J(B6) mice were treated with ENU and repro-duced ...Phenotype-driven is the name for an ap-proach used to study gene functions through mutagenesis, location and cloning of the mutant gene. In this study, 150 male C57BL/6J(B6) mice were treated with ENU and repro-duced a total offspring of 3860. Of these descendants, 210 exhibited mutation phenotypes by screening, and more than 10 of them are hereditable. Four kinds of mutant mice, named Wbct, W-1Bao, W-2Bao, and W-3Bao, showed dominant hereditary white spot mutation with partial albinism on their belly, distal limbs and tail terminal. To map these mutant genes, 39 microsatellites, equally distributed on the mouse genome and with difference between B6 and DBA/2J (D2), were selected to scan the genome after discrimination of the white spots in the F2 mice [(B6×D2)×D2]. It is found that, the log odds score (LODS) between W-1Bao and D5Mit168 is 0.56, and the LODS of W-1Bao and D5Mit352 is 4.47. With the gradual application of microsatellites D5Mit290, D5Mit312, D5Mit308 and D5Mit356 that are close to the mutant gene, and the number of F2 mice going up to 537, the mutant W-1Bao is located between D5Mit356 and D5Mit308 on chro-mosome 5, about 42.19 cM from the centromere. In the same way, W-2Bao and W-3Bao are mapped nearby W-1Bao, and Wbct is located on chromosome 1, about 41.6 cM from the centro-mere. After searching for the mouse genome database (MGD) and performing a one-by-one study of all genes located on chromosome subregion, it is believed that the kit gene is an excellent candidate for the white spot mutations of W-1Bao, W-2Bao and W-3Bao.展开更多
Chinese soft-shelled turtles （Trionyx sinens） in culture farms using an artificial warming system in Zhejiang, China, often show typical signs of white-spot disease such as white spots on their bodies, skin lesions,...Chinese soft-shelled turtles （Trionyx sinens） in culture farms using an artificial warming system in Zhejiang, China, often show typical signs of white-spot disease such as white spots on their bodies, skin lesions, anorexia and eventually death. The sick turtles were mostly 5-80 g in weight. A suspected fungal pathogen was isolated from the sick turtles and verified as Paecilomyces lilacinus by sequence analysis of the internal transcribed spacer （ITS） of its ribosomal DNA （rDNA）. Detailed morphological examinations were also conducted to confirm the white-spot disease.展开更多
In this study the kit gene was amplified from total brain RNA of W-3Bao heterozygote and normal C57BL/6 mice by RT-PCR followed by sequencing.To validate the mutation found in mRNA,the corresponding genome sequence wa...In this study the kit gene was amplified from total brain RNA of W-3Bao heterozygote and normal C57BL/6 mice by RT-PCR followed by sequencing.To validate the mutation found in mRNA,the corresponding genome sequence was amplified and sequenced.The results showed that there was a T-to-A transversion at the 441st nucleotide in W-3Bao ORF.The mutation introduces a pre-termination codon of amino acid and causes the phenotype of dominant white spot.展开更多
文摘Phenotype-driven is the name for an ap-proach used to study gene functions through mutagenesis, location and cloning of the mutant gene. In this study, 150 male C57BL/6J(B6) mice were treated with ENU and repro-duced a total offspring of 3860. Of these descendants, 210 exhibited mutation phenotypes by screening, and more than 10 of them are hereditable. Four kinds of mutant mice, named Wbct, W-1Bao, W-2Bao, and W-3Bao, showed dominant hereditary white spot mutation with partial albinism on their belly, distal limbs and tail terminal. To map these mutant genes, 39 microsatellites, equally distributed on the mouse genome and with difference between B6 and DBA/2J (D2), were selected to scan the genome after discrimination of the white spots in the F2 mice [(B6×D2)×D2]. It is found that, the log odds score (LODS) between W-1Bao and D5Mit168 is 0.56, and the LODS of W-1Bao and D5Mit352 is 4.47. With the gradual application of microsatellites D5Mit290, D5Mit312, D5Mit308 and D5Mit356 that are close to the mutant gene, and the number of F2 mice going up to 537, the mutant W-1Bao is located between D5Mit356 and D5Mit308 on chro-mosome 5, about 42.19 cM from the centromere. In the same way, W-2Bao and W-3Bao are mapped nearby W-1Bao, and Wbct is located on chromosome 1, about 41.6 cM from the centro-mere. After searching for the mouse genome database (MGD) and performing a one-by-one study of all genes located on chromosome subregion, it is believed that the kit gene is an excellent candidate for the white spot mutations of W-1Bao, W-2Bao and W-3Bao.
基金the Science and Technology Department of Zhejiang Province, China (No. 2004C26026)the Science and Technology Department of Hangzhou City, China (No. 20051322 B33)
文摘Chinese soft-shelled turtles （Trionyx sinens） in culture farms using an artificial warming system in Zhejiang, China, often show typical signs of white-spot disease such as white spots on their bodies, skin lesions, anorexia and eventually death. The sick turtles were mostly 5-80 g in weight. A suspected fungal pathogen was isolated from the sick turtles and verified as Paecilomyces lilacinus by sequence analysis of the internal transcribed spacer （ITS） of its ribosomal DNA （rDNA）. Detailed morphological examinations were also conducted to confirm the white-spot disease.
文摘In this study the kit gene was amplified from total brain RNA of W-3Bao heterozygote and normal C57BL/6 mice by RT-PCR followed by sequencing.To validate the mutation found in mRNA,the corresponding genome sequence was amplified and sequenced.The results showed that there was a T-to-A transversion at the 441st nucleotide in W-3Bao ORF.The mutation introduces a pre-termination codon of amino acid and causes the phenotype of dominant white spot.
