利用精子介导法建立<em><em>hHT/hDAF</em></em>双基因转移小鼠模型

doi:10.7668/hbnxb.2010.03.010

华北农学报 ›› 2010, Vol. 25 ›› Issue (3): 43-46. doi: 10.7668/hbnxb.2010.03.010

所属专题： 生物技术；

利用精子介导法建立hHT/hDAF双基因转移小鼠模型

郭战军¹, 马毅², 李三华³, 韩瑞发¹, 李胜芝¹, 孙光¹, 乔宝民¹, 马腾骧¹, 陈永福⁴

1. 天津市泌尿外科研究所, 天津 300211;
2. 天津市奶牛发展中心, 天津 300384;
3. 遵义医学院, 中心实验室, 贵州遵义 563003;
4. 中国农业大学, 生物学院, 北京 100094

收稿日期:2009-12-09 出版日期:2010-06-28
通讯作者: 乔宝民(1967－)，男，天津人，主任医师，博士，主要从事泌尿外科临床及异种移植基础研究。
作者简介:郭战军(1971-),男,山东淄博人,主治医师,在读博士,主要从事泌尿外科研究.
基金资助:
天津市自然科学基金(07JCYBJC1070008JCYBJC04500);贵州省科学技术基金([2009]2173)

Establishment of Double-gene Transgenic Mouse Model for hHT and hDAF through SMGT

GUO Zhan-jun¹, MA Yi², LI San-hua³, HAN Rui-fa¹, LI Sheng-zhi¹, SUN Guang¹, QIAO Bao-min¹, MA Teng-xiang¹, CHEN Yong-fu⁴

1. Tianjin Institute of Urological Surgery, Tianjin 300211, China;
2. Tianjin Dairy Cattle Development Center, Tianjin 300384, China;
3. Central Lab of Zunyi Medical College, Zunyi 563003, China;
4. College of Biology, China Agricultural University, Beijing 100094, China

Received:2009-12-09 Published:2010-06-28

摘要/Abstract

摘要： 为探讨超急排斥反应(Hyperacute rejection,HAR)及延迟性异种排斥反应(Delayed xenograft rejection,DXR)的分子调控机制,采用优化的精子介导法(Sperm-mediated gene transfer,SMGT)对建立hHT/hDAF双基因转移小鼠模型进行了研究。结果表明,应用树枝状聚合物(Dendrimers)优化SMGT后,2细胞胚GFP+RFP的阳性率达到4.51%,同对照组相比差异极显著(P<0.01)。选取标记基因GFP+RFP双阳性的胚胎进行移植后获得36只仔鼠,PCR及Southern Blot检测结果表明,17只小鼠PCR结果呈hHT+hDAF双阳性,其中5只小鼠Southern Blot结果呈hHT+hDAF双阳性。上述结果表明,成功建立了hHT/hDAF双基因转移小鼠模型。

关键词: 精子介导法, 双基因, hHT, hDAF, 转基因, 小鼠

Abstract: To explore the molecular mechanism of hyperacute rejection( HAR) and delayed xenograft rejection ( DXR), double-gene transgenic mouse model expressing Human alpha1,2-fucosyltransferase( hHT) and human-decay accelerating factor( hDAF) was established using sperm-mediated gene transfer optimized by dendrimers( SMGT).The positive rate of PCR for GFP + RFP in 2-cell embryos was 4.51% after the SMGT was optimized,which was significantly higher than that of the control group( P<0.01).Embryos expressing both GFP and RFP were got were transplanted to the pseudopregnant female mice.36 offspring were got.The founder transgenic mice( G0 mice) were identified with PCR and southern blot.The results of PCR showed that genomes from 17 mice were integrated by both hHT and hDAF.The results of southern blot for 5 mice showed a positive signal of hHT + hDAF.These results indicated that G0 double-gene transgenic mouse model for hHT and hDAF was established．

Key words: SMGT, Double-gene, hHT, hDAF, Transgenic, Mouse

中图分类号:

Q789

郭战军, 马毅, 李三华, 韩瑞发, 李胜芝, 孙光, 乔宝民, 马腾骧, 陈永福. 利用精子介导法建立hHT/hDAF双基因转移小鼠模型[J]. 华北农学报, 2010, 25(3): 43-46. doi: 10.7668/hbnxb.2010.03.010.

GUO Zhan-jun, MA Yi, LI San-hua, HAN Rui-fa, LI Sheng-zhi, SUN Guang, QIAO Bao-min, MA Teng-xiang, CHEN Yong-fu. Establishment of Double-gene Transgenic Mouse Model for hHT and hDAF through SMGT[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(3): 43-46. doi: 10.7668/hbnxb.2010.03.010.

http://www.hbnxb.net/CN/Y2010/V25/I3/43

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利用精子介导法建立hHT/hDAF双基因转移小鼠模型

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利用精子介导法建立hHT/hDAF双基因转移小鼠模型

Establishment of Double-gene Transgenic Mouse Model for hHT and hDAF through SMGT

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