以流式细胞仪分离小麂(Muntiacus reevesi)Y染色体和黑麂(Muntiacus crinifrons)Y1,Y2,X+4和1号染色体,利用DOP-PCR技术富集了分离的各单条染色体。然后,将小麂的Y染色体的DOP-PCR产物经Cy3标记后直接作为涂染探针, 应用染色体涂染技术...以流式细胞仪分离小麂(Muntiacus reevesi)Y染色体和黑麂(Muntiacus crinifrons)Y1,Y2,X+4和1号染色体,利用DOP-PCR技术富集了分离的各单条染色体。然后,将小麂的Y染色体的DOP-PCR产物经Cy3标记后直接作为涂染探针, 应用染色体涂染技术与雌雄黑麂的核型标本进行杂交,确认了黑麂真正的Y染色体为Y2染色体。再以黑麂的Y1,Y2,X+4和1号染色体的DOP-PCR产物为模板,用人的特异性的 SRY（sex determining region of the Y chromosome ） 基因引物对其进行扩增,结果表明黑麂只有Y2染色体出现了SRY扩增片段。然后扩增产物克隆和测序,比较它与人的同源性,初步把黑麂的Sry基因定位在Y2染色体上。最后提取雄性黑麂的基因组DNA,并用同一对引物对其进行扩增,亦得到Sry基因的片段,对此扩增片段进行克隆,测序,结果表明其与Y2染色体得到的Sry基因片段完全一样，与人SRY基因的同源性均为83%。展开更多
Ty1/copia-like sequences were amplified from mung bean(Vigna radiata(L.)Wilczek)genomic DNA,by PCR with degenerate oligonucleotide primers corresponding to highly conserved domains in the Ty1/copia-like retrotransposo...Ty1/copia-like sequences were amplified from mung bean(Vigna radiata(L.)Wilczek)genomic DNA,by PCR with degenerate oligonucleotide primers corresponding to highly conserved domains in the Ty1/copia-like retrotransposons.PCR fragments of roughly 270 bp were isolated and cloned,and forty clones were sequenced.Thirty-six of the forty clones had unique nucleotide sequences,and eighteen clones had a frameshift,a stop codon,or both.Alignment of the nucleotide sequences indicated that these clones,denoted Tvr,fell into nine subgroups and nine ungrouped sequences.The nucleotide sequence similarity between these elements ranged from 8%to 100%,which indicates high level of sequence heterogeneity among these clones.A phylogenetic analysis comparing these clones with corresponding sequences from other plant species showed that some of the Tvr clones are more closely related to Ty1/copia-like retrotransposons from other species than to other Tvr clones.Dot blot analysis revealed that Ty1/copia-like retrotransposons comprise about 9.3%of the mung bean genome.展开更多
研究利用玻璃针显微分离了巴马小型猪外周血淋巴细胞有丝分裂中期的1号染色体，然后将显微分离的染色体进行简并寡核苷酸引物PCR（DOP—PCR）扩增，通过PCR技术和荧光原位杂交（Florescence in situ hybridization，FISH）技术对扩增产...研究利用玻璃针显微分离了巴马小型猪外周血淋巴细胞有丝分裂中期的1号染色体，然后将显微分离的染色体进行简并寡核苷酸引物PCR（DOP—PCR）扩增，通过PCR技术和荧光原位杂交（Florescence in situ hybridization，FISH）技术对扩增产物来源进行鉴定后，将DOP—PCR产物与pMD18-T载体连接、转化以构建1号染色体微克隆文库。结果表明：DOP—PCR扩增产物与猪的1号染色体DNA同源，并且得到了插入片段为100～600bp的巴马小型猪的1号染色体微克隆DNA文库。展开更多
文摘以流式细胞仪分离小麂(Muntiacus reevesi)Y染色体和黑麂(Muntiacus crinifrons)Y1,Y2,X+4和1号染色体,利用DOP-PCR技术富集了分离的各单条染色体。然后,将小麂的Y染色体的DOP-PCR产物经Cy3标记后直接作为涂染探针, 应用染色体涂染技术与雌雄黑麂的核型标本进行杂交,确认了黑麂真正的Y染色体为Y2染色体。再以黑麂的Y1,Y2,X+4和1号染色体的DOP-PCR产物为模板,用人的特异性的 SRY（sex determining region of the Y chromosome ） 基因引物对其进行扩增,结果表明黑麂只有Y2染色体出现了SRY扩增片段。然后扩增产物克隆和测序,比较它与人的同源性,初步把黑麂的Sry基因定位在Y2染色体上。最后提取雄性黑麂的基因组DNA,并用同一对引物对其进行扩增,亦得到Sry基因的片段,对此扩增片段进行克隆,测序,结果表明其与Y2染色体得到的Sry基因片段完全一样，与人SRY基因的同源性均为83%。
文摘Ty1/copia-like sequences were amplified from mung bean(Vigna radiata(L.)Wilczek)genomic DNA,by PCR with degenerate oligonucleotide primers corresponding to highly conserved domains in the Ty1/copia-like retrotransposons.PCR fragments of roughly 270 bp were isolated and cloned,and forty clones were sequenced.Thirty-six of the forty clones had unique nucleotide sequences,and eighteen clones had a frameshift,a stop codon,or both.Alignment of the nucleotide sequences indicated that these clones,denoted Tvr,fell into nine subgroups and nine ungrouped sequences.The nucleotide sequence similarity between these elements ranged from 8%to 100%,which indicates high level of sequence heterogeneity among these clones.A phylogenetic analysis comparing these clones with corresponding sequences from other plant species showed that some of the Tvr clones are more closely related to Ty1/copia-like retrotransposons from other species than to other Tvr clones.Dot blot analysis revealed that Ty1/copia-like retrotransposons comprise about 9.3%of the mung bean genome.
文摘研究利用玻璃针显微分离了巴马小型猪外周血淋巴细胞有丝分裂中期的1号染色体，然后将显微分离的染色体进行简并寡核苷酸引物PCR（DOP—PCR）扩增，通过PCR技术和荧光原位杂交（Florescence in situ hybridization，FISH）技术对扩增产物来源进行鉴定后，将DOP—PCR产物与pMD18-T载体连接、转化以构建1号染色体微克隆文库。结果表明：DOP—PCR扩增产物与猪的1号染色体DNA同源，并且得到了插入片段为100～600bp的巴马小型猪的1号染色体微克隆DNA文库。
