The flow cytometer was used to compare the DNA content in the blood cells and sperm in the new type of tetraploid (G 1×4n) and in red crucian carp used as the control. The new type of tetraploid (G 1×4n) was...The flow cytometer was used to compare the DNA content in the blood cells and sperm in the new type of tetraploid (G 1×4n) and in red crucian carp used as the control. The new type of tetraploid (G 1×4n) was produced by mating the diploid female gynogenetic progeny (first generation,G 1) of the tetraploid hybrid of red crucain carp × common carp, with the male of the tetraploid hybrid. Chromosome spreads from the kidney tissue in the new type of tetraploid were also examined. The results from the flow cytometer indicated that the mean DNA content of the diploid sperm produced by the new type of tetraploid was twice that of the haploid sperm generated by the diploid red crucian carp, and the mean DNA content of the blood cells in the tetraploid was twice that in red crucian carp. Examination of chromosome spreads from the kidney tissue also indicated that the examined samples of the G 1×4n were tetraploids with 200 chromosomes, which was in agreement with the results of the DNA content measurements from the sperm and the blood cells. The present study proved that the new type of tetraploids was able to produce normal diploid sperm, which fertilized diploid eggs from the new type of tetraploid to produce viable offspring. The flow cytometer was proved to be one of the accurate, secure and simple methods to distinguish the different ploidy fish in blood cells and sperm by measuring their DNA content .展开更多
A Yunnan pony ear marginal tissue fibroblast cell line (NYPEM 2/2) was successfully established using the explant of the ear marginal tissue and then trypsinization the cells from the outgrowth. Observations on cell m...A Yunnan pony ear marginal tissue fibroblast cell line (NYPEM 2/2) was successfully established using the explant of the ear marginal tissue and then trypsinization the cells from the outgrowth. Observations on cell morphology and dynamic growth, analysis of karyotype and isoenzymes of lactate dehydrogenase and malate dehydrogenase were carried out. The expression of recombinant green fluorescence protein in the cells were also undertaken. The results showed that the population doubling time (PDT) of the cells was 24 h; the frequency of cell chromosome number to be 2n=64 was 92.9%; the banding patterns of the isozymes of the two enzymes had significant difference between the Yunnan pony ear marginal fibroblast cell line and the fibroblast cell lines of PEM 2/2, MSHEM 2/2 and BLCHE 2/2 derived from the Picdmont bovine ear, Mongolian ovine ear and Beijing local chicken embryo respectively. Tests for the contamination from bacteria, fungi or mycoplasma were negative; the transfection efficiency for the recombinant plasmid was 32.3%. This newly established cell line make the Yunnan pony breed, a national important genetic resource preserved at cell level, as well as will provide an effective experimental material for genetic studies on the Yunnan pony.展开更多
基金国家自然科学基金项目 (No 30170733 No 30330480)教育部跨世纪优秀人才培养基金 (No 200248) 项目资助~~
文摘The flow cytometer was used to compare the DNA content in the blood cells and sperm in the new type of tetraploid (G 1×4n) and in red crucian carp used as the control. The new type of tetraploid (G 1×4n) was produced by mating the diploid female gynogenetic progeny (first generation,G 1) of the tetraploid hybrid of red crucain carp × common carp, with the male of the tetraploid hybrid. Chromosome spreads from the kidney tissue in the new type of tetraploid were also examined. The results from the flow cytometer indicated that the mean DNA content of the diploid sperm produced by the new type of tetraploid was twice that of the haploid sperm generated by the diploid red crucian carp, and the mean DNA content of the blood cells in the tetraploid was twice that in red crucian carp. Examination of chromosome spreads from the kidney tissue also indicated that the examined samples of the G 1×4n were tetraploids with 200 chromosomes, which was in agreement with the results of the DNA content measurements from the sperm and the blood cells. The present study proved that the new type of tetraploids was able to produce normal diploid sperm, which fertilized diploid eggs from the new type of tetraploid to produce viable offspring. The flow cytometer was proved to be one of the accurate, secure and simple methods to distinguish the different ploidy fish in blood cells and sperm by measuring their DNA content .
基金国家基础性工作重大专项 (No . 2 0 0 1DEA10 0 0 6) 国家自然科学基金 (No . 3 0 10 0 13 2 )资助~~
文摘A Yunnan pony ear marginal tissue fibroblast cell line (NYPEM 2/2) was successfully established using the explant of the ear marginal tissue and then trypsinization the cells from the outgrowth. Observations on cell morphology and dynamic growth, analysis of karyotype and isoenzymes of lactate dehydrogenase and malate dehydrogenase were carried out. The expression of recombinant green fluorescence protein in the cells were also undertaken. The results showed that the population doubling time (PDT) of the cells was 24 h; the frequency of cell chromosome number to be 2n=64 was 92.9%; the banding patterns of the isozymes of the two enzymes had significant difference between the Yunnan pony ear marginal fibroblast cell line and the fibroblast cell lines of PEM 2/2, MSHEM 2/2 and BLCHE 2/2 derived from the Picdmont bovine ear, Mongolian ovine ear and Beijing local chicken embryo respectively. Tests for the contamination from bacteria, fungi or mycoplasma were negative; the transfection efficiency for the recombinant plasmid was 32.3%. This newly established cell line make the Yunnan pony breed, a national important genetic resource preserved at cell level, as well as will provide an effective experimental material for genetic studies on the Yunnan pony.
