| 何丹丹,邹修平,解栋梁,彭爱红,许兰珍,何永睿,陈善春.融合标记基因NPTⅡ∷GFP在柑橘遗传转化中筛选效率的研究[J].中国南方果树,2014,43(6): |
| 融合标记基因NPTⅡ∷GFP在柑橘遗传转化中筛选效率的研究 |
| Transformation efficiency of a fusion gene NPTII ::GFP in citrus |
| 投稿时间：2014-06-18 修订日期：2014-07-08 |
| DOI： |
| 中文关键词: nptⅡ∷GFP融合标记基因 柑橘遗传转化 转化效率 |
| 英文关键词:NPTⅡ∷GFP fusion gene Citrus transformation Transformation efficiency |
| 基金项目: |
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| 中文摘要: |
| 绿色荧光蛋白（GFP）基因和新霉素磷酸转移酶（NPTⅡ）基因被广泛应用于植物的遗传转化中。为了研究融合标记基因NPTⅡ::GFP在柑橘遗传转化中的筛选效率，本实验将CaMV35S组成型启动子驱动的nptⅡ∷GFP融合标记基因表达载体pNGM和由CaMV35S启动子分别驱动NPTⅡ、GFP两个基因的表达载体pNPTII-GF经农杆菌介导法转化锦橙上胚轴。GFP绿色荧光检测结果显示，GFP在转NPTⅡ∷GFP融合基因植株中强烈表达，荧光强度与非融合GFP转基因植株的表达水平相当。NPTⅡ∷GFP融合标记基因对锦橙的转化效率为10.8%，与非融合NPTⅡ基因的转化效率（11.0%）没有明显差异。研究结果表明，NPTⅡ∷GFP融合标记基因是一个有效的柑桔遗传转化筛选标记基因。 |
| 英文摘要: |
| Green fluorescent protein (GFP) gene and neomycin phosphoric acid transferase (NPT Ⅱ) gene have been widely used in plant genetic transformation. In order to investigate the genetic transformation efficiency of fusion gene NPTⅡ::GFP in citrus, we constructed Agrobactirum-mediated genetic transformation of Jin Cheng (Citrus sinensis) with the pNGM vector containing the NPTⅡ:GFP fusion gene under the control of CaMV 35S promoter, using the pNPTⅡ-GF vector containing both NPTⅡ gene and GFP gene as a control. Results showed that GFP expressed strongly in the fusion gene NPTⅡ::GFP transfromed plants. The transformation efficiency of NPTⅡ::GFP fusion gene was 10.8%, comparable with that (11.0%) of NPTⅡ gene. The results revealed that the fusion gene NPTⅡ∷GFP is an effective selectable marker gene of citrus genetic transformation. |
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