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国家自然科学基金(31000546)

作品数:3 被引量:10H指数:2
相关作者:魏泓葛亮商海涛肖榕郑小波更多>>
相关机构:第三军医大学西南大学更多>>
发文基金:国家自然科学基金国家高技术研究发展计划国家科技支撑计划更多>>
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巴马香猪类固醇激素急性调节蛋白(StAR)基因真核表达载体的构建及鉴定
2013年
本试验通过RT-PCR扩增巴马香猪StAR基因,分析其序列的结构特点并构建真核表达载体,为StAR基因在猪睾丸间质细胞中的超量表达研究奠定基础。根据NCBI中猪StAR基因序列设计并合成引物,以巴马香猪睾丸组织总RNA为模板进行RT-PCR,将所得目的基因与pEGFP-C1进行双酶切后连接构建真核表达载体pEGFP-C1-StAR。结果显示,连接到pSURE-T克隆载体上的目的片段测序结果为858碱基组成,编码含有285个氨基酸残基的蛋白质,与NCBI报道的StAR基因比对,核酸同源性在99.3%以上,并成功构建了重组载体pEGFP-C1-StAR。
吴群魏泓肖榕卿利娟葛亮李婧商海涛郑小波刘宇
关键词:真核表达载体
Nucleus Transfer Efficiency of Ear Fibroblast Cells Isolated from Bama Miniature Pigs at Various Ages被引量:3
2015年
Somatic cell nucleus transfer(SCNT) has been considered the most effective method for conserving endangered animals and expanding the quantity of adult animal models. Bama miniature pigs are genetically stable and share similar biological features to humans. These pigs have been used to establish animal models for human diseases, and for many other applications. However, there is a paucity of studies on the effect of ear fibroblasts derived from different age of adult Bama miniature pigs on nucleus transfer(NT). The present study examined the NT efficiency of ear fibroblasts from fetal, newborn, 1-, 2-, 4-, 6-, 12-month-old miniature pigs by using trypan blue staining, flow cytometry and NT technique, etc., and the cell biological function and SCNT efficiency were compared between groups. The results showed that ear fibroblasts grew well after passage in each group. Spindle-shaped cells initially predominated, and gradually declined with increase of culture time and replaced by polygonal cells. Irregular cell growth occurred in the 2-month-old group and the elder groups. The growth curves of the ear fibroblasts were "S-shaped" in different age groups. The cell proliferation of postnatal ear fibroblasts, especially those from 2-, 4-, 6-, 12-month-old miniature pigs was significantly different from that of fetus ear fibroblasts(P<0.05 or P<0.01). Two-month- and 4-month-old ear fibroblasts had a significantly higher proportion of G1 stage cells(85% to 91%) than those at 6 and 12 months(66% to 74%, P<0.01). The blastocyst rate of reconstructed embryos originating from newborn, 1-, 2-, 4-month-old donor pigs was 6.06% to 7.69% with no significant difference from that in fetus fibroblast group(8.06%). It was concluded that <4-month-old adult Bama miniature pigs represent a better donor cell resource than elder pigs.
王庆华彭云蔡信永万萌刘宇魏泓
关键词:巴马小型猪胎儿成纤维细胞体细胞核移植
仔猪睾丸间质细胞培养方法的比较被引量:7
2013年
为了对比仔猪睾丸间质细胞不同培养方法的培养效果。运用过筛培养法、不过筛培养法,组织块培养法3种方法对仔猪睾丸间质细胞进行体外分离和培养,并对其进行细胞各项指标的比较。结果显示,不过筛培养法培养的细胞活力、纯度、细胞功能都较好,培养时间也短;过筛培养法培养的细胞活力较低;组织块培养法培养的细胞纯度较低。但在其他细胞指标上,后两种方法又各自有优点。结果3种培养方法都能简单有效的培养出状态良好的仔猪睾丸间质细胞,但其在纯度,活力,培养时间等方面有所差别。
卿利娟魏泓李婧肖榕吴群葛亮商海涛郑小波刘宇
关键词:睾丸间质细胞纯度
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