Polymorphic microsatellites are among the versatile genetic markers in molecular ecology studies. In contrast to de novo isolation of microsatellites from target species, cross-species amplification is a costeffective...Polymorphic microsatellites are among the versatile genetic markers in molecular ecology studies. In contrast to de novo isolation of microsatellites from target species, cross-species amplification is a costeffective approach for a fast development of microsatellite markers from closely related taxa. In our study, we crossamplified a panel of polymorphic microsatellite markers for the Blue Eared Pheasant (Crossoptilon auritum), a species endemic to China of considerable conservation concern. We obtained 11 polymorphic microsatellite markers selected from 112 candidate loci, originally isolated from other Galliforme species. This panel of makers has shown moderate to high levels of polymorphism and include a Z-chromosomal linkage locus. We carried out preliminary analyses of parentage among captive individuals with a known pedigree using this new panel of microsatellites. Our results suggest that the high utility of these markers may be powerful tools for studies in conservation genetics of eared-pheasants and other endangered Galliforme species.展开更多
We applied a resistance split-fusion strategy to increase the in vivo direct cloning efficiency mediated by Red recombination. The cat cassette was divided into two parts： cma （which has a homologous sequence with ...We applied a resistance split-fusion strategy to increase the in vivo direct cloning efficiency mediated by Red recombination. The cat cassette was divided into two parts： cma （which has a homologous sequence with cmb） and cmb, each of which has no resistance separately unless the two parts are fused together. The crab sequence was integrated into one flank of a target clon- ing region in the chromosome, and a linear vector containing the cma sequence was electroporated into the cells to directly capture the target region. Based on this strategy, we successfully cloned an approximately 48 kb DNA fragment from the E. coli DH1-Z chromosome with a positive frequency of approximately 80%. Combined with double-strand breakage-stimulated homologous recombination, we applied this strategy to successfully replace the corresponding region of the E. coli DH36 chromosome and knock out four non-essential genomic regions in one step. This strategy could provide a powerful tool for the heterologous expression of microbial natural product biosynthetic pathways for genome assembly and for the functional study of DNA sequences dozens of kilobases in length.展开更多
基金supported by the National Key Project of the Scientific and Technical Supporting Programs Funded by the Ministry of Science and Technology of China(2012BAC01B06)the National Natural Science Foundation of China (No. 30570234)
文摘Polymorphic microsatellites are among the versatile genetic markers in molecular ecology studies. In contrast to de novo isolation of microsatellites from target species, cross-species amplification is a costeffective approach for a fast development of microsatellite markers from closely related taxa. In our study, we crossamplified a panel of polymorphic microsatellite markers for the Blue Eared Pheasant (Crossoptilon auritum), a species endemic to China of considerable conservation concern. We obtained 11 polymorphic microsatellite markers selected from 112 candidate loci, originally isolated from other Galliforme species. This panel of makers has shown moderate to high levels of polymorphism and include a Z-chromosomal linkage locus. We carried out preliminary analyses of parentage among captive individuals with a known pedigree using this new panel of microsatellites. Our results suggest that the high utility of these markers may be powerful tools for studies in conservation genetics of eared-pheasants and other endangered Galliforme species.
基金supported by the National Natural Science Foundation of China(81373286)National Basic Research Program of China(2011CBA00800)
文摘We applied a resistance split-fusion strategy to increase the in vivo direct cloning efficiency mediated by Red recombination. The cat cassette was divided into two parts： cma （which has a homologous sequence with cmb） and cmb, each of which has no resistance separately unless the two parts are fused together. The crab sequence was integrated into one flank of a target clon- ing region in the chromosome, and a linear vector containing the cma sequence was electroporated into the cells to directly capture the target region. Based on this strategy, we successfully cloned an approximately 48 kb DNA fragment from the E. coli DH1-Z chromosome with a positive frequency of approximately 80%. Combined with double-strand breakage-stimulated homologous recombination, we applied this strategy to successfully replace the corresponding region of the E. coli DH36 chromosome and knock out four non-essential genomic regions in one step. This strategy could provide a powerful tool for the heterologous expression of microbial natural product biosynthetic pathways for genome assembly and for the functional study of DNA sequences dozens of kilobases in length.
