河套蜜瓜花粉管通道法转化hALR基因  

Transforming hALR Gene into Cucumis melo L. cv Hetao via Pollen-tube Pathway

作　　者：张立全[1] 哈斯阿古拉[1] 扈廷茂[1] 刘明秋[1] 

机构地区：[1]内蒙古大学生命科学学院生物工程中心内蒙古自治区牧草与特色作物生物技术重点实验室,呼和浩特010021

出　　处：《生物技术通报》2009年第3期70-73,共4页Biotechnology Bulletin

基　　金：内蒙古自然科学基金项目(200208020307)

摘　　要：为利用河套蜜瓜为生物反应器生产可用于治疗肝病的蛋白因子,以人胎肝mRNA为模板,经RT-PCR扩增获得了471 bp cDNA片段。克隆到pMD18-T载体,命名为pMD-ALR,测序分析结果与GenBank中报道的hALR编码序列完全相同。酶切回收hALR片段,插入到含有甜瓜果实黄瓜素(Cucumisin)基因启动子的果实特异性表达载体pPZP-CGN中,构建了hALR甜瓜果实特异性表达载体pPZP-CAN。花粉管通道法转化试验大田自花授粉花63朵,收获T0代果实27颗,结实率为42.9%。提取T0代种子无菌苗DNA,经PCR、PCR-southern和斑点杂交检测有13株呈阳性。In order to use Cucumis melo L. cv Hetao as a bioreactor for production of human augmenter of liver regeneration （hALR） protein,471 bp cDNA hALR was amplified from the human fetal liver mRNA by RT-PCR with two artificially synthesized primers,then ligated into vector pMD18-T to construct pMD-ALR. The sequence analysis of pMD-ALR showed that the sequence was completely identical with that of bALK eDNA in GenBank. The fruit-specific expression vector pPZP-CAN was constructed by inserting bALK which was obtained from pMD-ALR by Hind Ⅲ and Sac Ⅰ restrictive digesting,into plant expression vector pPZP-CGN which contains Cncumis me/o L. cv cucumisin gene promoter. 63 hand-pollinating flowers were obtained via pollen-tube pathway transformation method in testing-farm,by which 27 fruits were achieved with 42.9 percentage of fruit-set. The DNA from To seedings were analyzed by PCR, PCR-southern and Dot boltting analysis and 13 positive were obtained.

关 键 词：人肝再生增强因子 果实特异性 花粉管通道法 河套蜜瓜 

分 类 号：S652[农业科学—果树学] Q943[农业科学—园艺学]