【作者】 周向梅； 马月辉； 关伟军； 赵德明；

【Author】 ZHOU Xiang-mei, MA Yue-hui, GUAN Wei-jun and ZHAO De-ming(Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100094,P.R. China. College of Veterinary Medicine, China Agricultural University, Beijing 100094,P.R. China.)

【机构】 中国农业科学院畜牧研究所； 中国农业大学动物医学院；

【摘要】 本试验采用组织块直接培养法,对德保矮马耳缘组织进行原代和继代培养,成功地建立了成纤维细胞系。试验中对培养细胞了进行形态学、细胞生长动态学和重组绿色荧光蛋白在培养细胞中表达的观察,以及核型和乳酸脱氢酶、苹果酸脱氢酶两种酶的同工酶分析。结果表明细胞群体倍增时间(PDT)为48小时;细血染色体众数2n=64的细胞数占92.9%;两种酶的同工酶电泳图谱与本室其它畜禽品种细胞系有明显差别;细菌、真菌、病毒、支原体检测呈阴性;细胞转染绿色荧光蛋白的效率较高,达32.3%。该细胞系的建立,使德保矮马这一国家重要遗传资源在细胞水平上得以保存下来,也为进一步基因组和体细胞克隆等研究提供了理想的生物材料。更多还原

【Abstract】 A Debao pony ear marginal tissue fibroblast cell line (NDPE 2/2) was successfully established using the primary explant technique. Observations on morphology, dynamic growth and expression of recombinant green fluorescence protein in the cells were undertaken, and analysis of karyotype, isoenzymes of lactate dehydrogenase and malate dehydrogenase were carried out. The results showed that the population doubling tune (PDT) of cells was 48 h; the frequency of cell chromosome number to be 2n = 64 was 92.9%; the banding patterns of the isoenzymes of the two enzymes had significant difference between the NDPE and other cell Unes in our laboratory; tests for microbial contamination from bacteria, fungi or mycoplasma were negative; transfer efficiency for the plasmid was rather high of 30%. This newly established cell line will ensure the Debao pony breed, a nationally important genetic resource preserved in cell level, and will provide an ideal biological material for the genetic studies on Debao pony as well.更多还原