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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2013, Vol. 33 Issue (6): 131-137    
农业生物技术专栏     
转IL-4基因大白菜后代的分子检测及遗传分析
金晓霞, 高亚, 于丽杰
哈尔滨师范大学生命科学与技术学院 哈尔滨 150025
Molecular Detection and Genetic Analysis of Chinese Cabbage Progeny with the IL-4
JIN Xiao-xia, GAO Ya, YU Li-jie
College of Life Science and Technology, Harbin Normal University, Harbin 150025, China
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摘要：

采用PPT抗性筛选、PCR扩增、PCR-Southern杂交、ELISA、Western杂交等方法,对花粉管通道法介导IL-4 基因转化大白菜的后代进行了分子生物学检测,同时结合统计分析手段研究了IL-4 基因在转基因大白菜后代中的遗传规律。结果表明:(1)PPT抗性的转基因大白菜,T4代植株PPT抗性的阳性率为33.43%,IL-4 基因PCR扩增的阳性率为1.39%。IL-4 的分离情况不符合孟德尔分离定律。(2)对T4代5个株系的14个PCR阳性株进行ELISA检测,其中有7份样品为阳性,IL-4表达量约为326.87~1233.13 ng/g FW,同一株系内各株间IL-4的表达量差异显著。(3)Western杂交进一步证实IL-4在转基因大白菜后代中能正常表达。PCR-southern杂交表明IL-4 基因已整合到大白菜基因组中,且在转基因后代植株中仍然存在。
关键词： 花粉管通道法;  大白菜;  白细胞介素4;  非孟德尔遗传    
Abstract:

IL-4 gene transformation mediated by pollen-tube pathway method of Chinese cabbage for molecular detection of descendants using PPT resistance screening, PCR amplification, PCR-Southern hybrid, ELISA, Western hybrid methods, combined with statistical analysis study IL-4 inheritance of genes in transgenic Chinese cabbage in future generations. Results indicated that: (1) PPT resistance in transgenic Chinese cabbage, PPT resistance positive rates of T4 generation was 33.43%, the positive rate of PCR amplification of IL-4 gene was 1.39%. IL-4 separation plant does not meet Mendel’s law of segregation. (2) ELISA test on 14 ones of PCR-positive strains of 5 strains in the T4 generation, 7 samples of which are positive, IL-4 expression was about 326.87~1233.13 ng/g FW, the ones of IL-4 expression within the same strains significance of difference. (3) Western confirmed IL-4 in transgenic Chinese cabbage hybrid offspring in the regular expression. It indicated that IL-4 gene has been incorporated into the Chinese cabbage in the genome with PCR-southern hybrid, and that gene still exists in transgenic plants’ future generations.
Key words: Pollen-tube pathway method    Chinese cabbage    Interleukin-4 (IL-4)    Non-Mendelian Inheritance
收稿日期: 2013-02-19 出版日期: 2013-06-25
ZTFLH:  Q819  
基金资助:

黑龙江省自然科学基金资助项目(C0014)
通讯作者: 于丽杰     E-mail: yulijie1961@126.com
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金晓霞
高亚
于丽杰

引用本文:

金晓霞, 高亚, 于丽杰. 转IL-4基因大白菜后代的分子检测及遗传分析[J]. 中国生物工程杂志, 2013, 33(6): 131-137.

JIN Xiao-xia, GAO Ya, YU Li-jie. Molecular Detection and Genetic Analysis of Chinese Cabbage Progeny with the IL-4. China Biotechnology, 2013, 33(6): 131-137.

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https://manu60.magtech.com.cn/biotech/CN/    或    https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I6/131

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