Transformation of vp1 Gene of Foot-and-Mouth Disease Virus in Legume Forage Lotus cornicu- latus L

WANG Bao-qin; WANG Xiao-long; ZHANG Yong-guang; PAN Li; WANG Wen-xiu; DONG Jin-jie

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October 2005

WANG Bao-qin, WANG Xiao-long, ZHANG Yong-guang, PAN Li, WANG Wen-xiu and DONG Jin-jie. Transformation of vp1 Gene of Foot-and-Mouth Disease Virus in Legume Forage Lotus cornicu- latus L[J]. Virologica Sinica, 2005, 20(5): 526-529.

Citation: WANG Bao-qin, WANG Xiao-long, ZHANG Yong-guang, PAN Li, WANG Wen-xiu, DONG Jin-jie. Transformation of vp1 Gene of Foot-and-Mouth Disease Virus in Legume Forage Lotus cornicu- latus L .VIROLOGICA SINICA, 2005, 20(5) : 526-529.

FMDV vp1基因在豆科牧草百脉根中的转化与表达

王宝琴 ^1,2 ,
王小龙 ¹ ,
张永光 ^2* ,
潘丽 ² ,
王文秀 ² ,
董金杰 ²

1.
1.江苏南京南京农业大学动物医学院 2.中国农业科学院兰州兽医研究所甘肃兰州

摘要

通过根癌农杆菌介导法,将FMDV阿克苏(Akesu/O/58)株结构基因vp1转化豆科牧草百脉根子叶和子叶柄,其愈伤、芽和生根等过程经50 mg/L Kan筛选后,获得Kan抗性百脉根植株。对抗性植株进行vp1基因的PCR、RT-PCR检测和VP1蛋白的Western-blotting杂交。结果表明:vp1基因转入百脉根中,检测有转录活性;目的蛋白获得了正确表达;扩繁和移栽后获得了批量转基因百脉根,为下一阶段的动物试验提供了实验材料。
- 口蹄疫病毒
- , 根癌农杆菌
- , vp1基因
- , 转化
- , 表达
- , 百脉根

Transformation of vp1 Gene of Foot-and-Mouth Disease Virus in Legume Forage Lotus cornicu- latus L

Abstract

The cotyledons and cotyledon petioles of legume forage Lotus corniculatus L.were transformed by Agrobacterium tumefaciens-mediated method with the binary expression vector pBinFMDV-VP1,which contained the vp1gene of FMDV strain Akesu/58 serotype O.Kanamycin-resistant plants were obtained after selected with 50 mg/L kanamycin.The vp1gene was analyzed through PCR with total DNA and through RT-PCR with total RNA from fresh leaves tissue of the kanamycin-resistant lines,the random samples from positive Lotus corniculatus L.lines were analyzed by Western-blotting.These results showed that the vp1 gene was transformed into Lotus corniculatus L.plants and had transcription activity;the VP1 protein was expressed correctly.The transgenic lines was transplanted to greenhouse,and further work is under way.
- Foot-and-mouth disease virus
- , Agrobacterium
- , Tumefaciens vp1gene
- , Transformation
- , Expression
- , Lotus corniculatus L.

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Transformation of vp1 Gene of Foot-and-Mouth Disease Virus in Legume Forage Lotus cornicu- latus L

Abstract: The cotyledons and cotyledon petioles of legume forage Lotus corniculatus L.were transformed by Agrobacterium tumefaciens-mediated method with the binary expression vector pBinFMDV-VP1,which contained the vp1gene of FMDV strain Akesu/58 serotype O.Kanamycin-resistant plants were obtained after selected with 50 mg/L kanamycin.The vp1gene was analyzed through PCR with total DNA and through RT-PCR with total RNA from fresh leaves tissue of the kanamycin-resistant lines,the random samples from positive Lotus corniculatus L.lines were analyzed by Western-blotting.These results showed that the vp1 gene was transformed into Lotus corniculatus L.plants and had transcription activity;the VP1 protein was expressed correctly.The transgenic lines was transplanted to greenhouse,and further work is under way.