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[prdm1基因敲除小鼠的繁殖及其基因型鉴定]

[Propagation of prdm1 gene knockout mouse and its genotype identification].

作者信息

Lu Xiao-Yun, Chen Chong, Pan Xiu-Ying, Zeng Ling-Yu, Li Zhen-Yu, Song Xu-Guang, Xu Kai-Lin

机构信息

Affiliated Hospital of Xuzhou Medical College, Jiangsu Province, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2012 Aug;20(4):985-8.

Abstract

This study was aimed to propagate and identify the prdm1 gene-knockout mice, so as to lay the foundation for studying Blimp-1 protein. Two kinds of transgenic homozygous mice with B6.prdm1(flox/flox) and B6.Lck-Cre were feed and propagated; after successful propagating, the first passage mice were obtained; after the first passage mice were copulated once again, the genotypes were obtained as follows: B6. prdm1(wild/wild). Lck-Cre, B6. prdm1(wild/wild), B6.prdm1(flox/flox). Lck-Cre, B6.prdm1(flox/wild). Lck-Cre, B6.prdm1(flox/flox), B6. prdm1(flox/wild). The genomic DNA of second passage mice was extracted, the Cre and loxp gene fragments were amplified by PCR, then the size of Cre and loxp genomic DNA were detected by agarose gel electrophoresis. The mice with B6.prdm1(flow/flox). Lek-Cre were used as conditionally prdm1-knockout mice, B6.prdm1(flox/wild). Lck-Cre mice, B6.prdm1(flox/flox) and B6 mice were used as controls. The spleen T lymphocytes and B lymphocytes were sorted by using magnetic beads, the blimp-1 target protein was identified by Western blot. The results showed that the two transgenic homozygous mice had the ability to reproduce, and the separation ratio of second passage mice generated from propagation of their offspring cach other meet Mendelian laws, and the prdm1 gene-knockout mice also could successfully obtained. It is concluded that the application of Cre-loxp system may successfully obtain plentiful prdm1 gene-knockout mice.

摘要

本研究旨在繁殖和鉴定prdm1基因敲除小鼠,为研究Blimp-1蛋白奠定基础。饲养并繁殖了两种转基因纯合小鼠,即B6.prdm1(flox/flox)和B6.Lck-Cre;繁殖成功后,获得了第一代小鼠;第一代小鼠再次交配后,得到如下基因型:B6. prdm1(野生/野生). Lck-Cre、B6. prdm1(野生/野生)、B6.prdm1(flox/flox). Lck-Cre、B6.prdm1(flox/野生). Lck-Cre、B6.prdm1(flox/flox)、B6. prdm1(flox/野生)。提取第二代小鼠的基因组DNA,通过PCR扩增Cre和loxp基因片段,然后用琼脂糖凝胶电泳检测Cre和loxp基因组DNA的大小。将B6.prdm1(flow/flox). Lek-Cre小鼠作为条件性prdm1基因敲除小鼠,B6.prdm1(flox/野生). Lck-Cre小鼠、B6.prdm1(flox/flox)小鼠和B6小鼠作为对照。用磁珠分选脾脏T淋巴细胞和B淋巴细胞,通过蛋白质免疫印迹法鉴定blimp-1靶蛋白。结果表明,两种转基因纯合小鼠具有繁殖能力,其后代相互繁殖产生的第二代小鼠的分离比例符合孟德尔定律,且prdm1基因敲除小鼠也能成功获得。结论是,Cre-loxp系统的应用可成功获得大量prdm1基因敲除小鼠。

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