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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2006, Vol. 26 Issue (0): 69-72    
专稿     
利用免疫细胞化学方法快速检测精子对外源DNA的结合能力
叶华虎 袁菊芳 温叶飞 周艳荣 熊福银 邓继先 刘彦
军事医学科学院生物工程研究所 军事医学科学院生物工程研究所 军事医学科学院生物工程研究所 军事医学科学院生物工程研究所 北京市农林科学院畜牧兽医研究所
Quickly evaluating the capability of sperm up-take exogenous DNA by immunocytochemistry
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摘要:

目的 建立评价精子结合外源DNA能力的简单、快速方法。方法 地高辛标记的DNA分子与山羊精子共同孵育,利用免疫细胞化学方法检测精子对外源DNA的结合能力以及外源DNA的作用特征,并将结果与传统的液体闪烁计数方法进行相关性分析。结果 结合外源DNA的阳性精子呈暗红色着染,阴性精子不着色,二者反差明显,易于辨认;同时,免疫细胞化学检测方法不仅能很好地反映精子对外源DNA的结合能力(阳性精子比例),而且能清楚显示外源DNA的作用过程和结合部位;此外,免疫细胞化学方法所得到的阳性结果与液闪法实验结果具有显著相关性(r =0.96)。结论 对于精子结合外源DNA能力的评价,免疫细胞化学方法具有安全、快速、准确等优点,而且该方法能更多地展示精子与DNA相互作用的某些特征。

关键词: 地高辛标记DNA免疫细胞化学精子转染液体闪烁计数    
Abstract:

Objective To establish a quick and simple method used for evaluating capability of sperm cells up-take exogenous DNA. Methods After co-cultured with digoxigenin (DIG) labeled DNA, frequencies of sperm cells carrying foreign DNA were tested by immunocytochemistry and compared with corresponding results from liquid scintillation counting. Results Sperm cells carrying exogenous DNA were dyed heavily red and control sperm cells did not. At the same time, the process and location of exogenous DNA interacted with sperm cells were also displayed. Furthermore, results came from immunocytochemistry had obviously relationship with that that from liquid scintillation counting, a classical method to evaluate sperm capability to take up foreign DNA (r = 0.96). Conclusions Imunocytochemistry is not only a safe, quickly and sensitive method in estimating the capability of sperm cells to take up exogenous DNA, but more information about interaction between sperm cells and exogenous DNA could also be revealed.

Key words: DIG labeled DNA    immunocytochemistry    sperm transfection    liquid scintillation counting
收稿日期: 2006-01-06 出版日期: 2006-06-15
通讯作者: 叶华虎   
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袁菊芳
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熊福银
温叶飞
叶华虎
刘彦

引用本文:

叶华虎,袁菊芳,温叶飞,周艳荣,熊福银,邓继先,刘彦. 利用免疫细胞化学方法快速检测精子对外源DNA的结合能力[J]. 中国生物工程杂志, 2006, 26(0): 69-72.

. Quickly evaluating the capability of sperm up-take exogenous DNA by immunocytochemistry. China Biotechnology, 2006, 26(0): 69-72.

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https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I0/69

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