| 引用本文: | 徐春香, 何勇强, 尉义明, 卢博彬, 胡桂兵, 陈厚彬.抗冻蛋白(afp)基因表达载体的构建及对香蕉胚性细胞悬浮系的转化[J].广西植物,2009,(5):664-668.[点击复制] |
| XU Chun-Xiang, HE Yong-Qiang, WEI Yi-Ming,
LU Bo-Bin, HU Gui-Bing, CHEN Hou-Bin.Construction of a plant expression vector containing antifreeze protein(afp)gene and it's transformation into banana(Musa AAA Cavendish)embryogenic cell suspension[J].Guihaia,2009,(5):664-668.[点击复制] |
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| 摘要: |
| 通过PCR从‘京都七寸人参'胡萝卜基因组DNA中扩增抗冻蛋白基因,测序结果表明该基因的核苷酸序列与从宁夏‘吴忠'胡萝卜中克隆的完全一致。先后将获得的胡萝卜afp基因克隆和亚克隆至pMD18-T和pBI121,构建植物表达载体pBI121-afp。通过冻融法将pBI121-afp导入根癌农杆菌EHA105中。以香蕉栽培品种‘北大矮蕉'的胚性细胞悬浮系为受体,采用农杆菌介导法将胡萝卜afp基因导入其中,然后在Kanamycin的选择压力下通过体细胞胚发生途径进行植株再生。共获得抗性再生植株9株,其中两株经PCR检测呈阳性,可初步确定目的基因已经整合到这两株转基因香蕉植株的基因组中。 |
| 关键词: 香蕉 胚性细胞悬浮系 根癌农杆菌介导法 遗传转化 |
| DOI： |
| 分类号:Q781 |
| 基金项目: |
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| Construction of a plant expression vector containing antifreeze protein(afp)gene and it's transformation into banana(Musa AAA Cavendish)embryogenic cell suspension |
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XU Chun-Xiang1, HE Yong-Qiang2, WEI Yi-Ming1,
LU Bo-Bin1, HU Gui-Bing1, CHEN Hou-Bin1
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1.College of Horticulture, South China Agricultural University, Guangzhou 510642;2.College of Life Science and Technology, Guangxi University, Nanning 530005
1.College of Horticulture, South China Agricultural University, Guangzhou 510642;
2.College of Life Science and Technology, Guangxi University, Nanning 530005
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| Abstract: |
| Antifreeze protein(afp)gene was amplified by polymerase chain reaction using genomic DNA extracted from the seedling of carrot cv.Jingdou Qicunrenshen. Sequencing result showed that there was no difference in nucleotide sequence between this afp gene and that cloned from‘Wuzhong'carrot in Ningxia. afp gene was subsequently cloned into intermediate vector pMD18-T and binary vector pBI121 to construct plant express vector pBI121-afp,which was transformed into Agrobacterium tumefaciens strain EHA105 by freeze-thaw method. Agrobacterium tumefaciens-mediated method was used to introduce afp gene into the embryogenic cell suspension of Musa AAA Cavendish cv.Beida Aijiao,followed by plant regeneration via embryogenesis. PCR analysis showed that 2 from 9 obtained regenerated plants resistant to kanamycin were positive,which possibly indicated that afp gene has been integrated into the genome of banana plants. |
| Key words: Banana embryogenic cell suspension Agrobacterium tumefaciens-mediated method genetic transformation |
