植物学报 ›› 2007, Vol. 24 ›› Issue (05): 609-613.

• 实验简报 • 上一篇    下一篇

口蹄疫抗原决定簇融合基因转化生菜及表达

邓小莉 常景玲   

  1. 1 平原大学应用生物系分子生物学实验室, 新乡453003; 2 河南科技学院生物工程系分子生物学重点实验室, 新乡 453003
  • 收稿日期:2007-01-12 修回日期:2007-04-05 出版日期:2007-09-01 发布日期:2007-09-01
  • 通讯作者: 邓小莉

Transformation and Expression of FMD Antigenic-determinant

Xiaoli Deng Jingling Chang   

  1. 1Laboratory of Molecular Biology, PingYuan University, Xinxiang 453003, China
  • Received:2007-01-12 Revised:2007-04-05 Online:2007-09-01 Published:2007-09-01
  • Contact: Xiaoli Deng

摘要: 以3-5天苗龄的散叶大速生生菜(Lactuca sativa var. capatata)无菌苗子叶为外植体, 通过根癌农杆菌介导, 将携带O型和A型口蹄疫抗原决定簇融合基因O21- O14 -A21-HBcAg导入生菜。研究结果表明, 含有20 mg.L-1潮霉素(Hyg)的S2 培养基(MS+1.5 mg.L-1 6-BA+0.2 mg.L-1 IAA+20 mg.L-1 Hyg +300 mg.L-1 Cb)为子叶外植体转化后诱导愈伤和芽再生的最适培养基, 经抗性筛选, 将抗性芽切下于S3 培养基(1/2MS+20 mg.L-1 Hyg)上诱导生根。通过PCR 和outhern杂交分析证明, 基因已经整合到生菜基因组中。RT-PCR检测初步表明, O21-O14 -A21-HBcAg基因可以在生菜中正常转录。

Abstract: The FMD antigenic-determinant fused gene (O21-O14 -A21-HbcAg) was introduced into lettuce (Lactuca sativa var. capatata) by co-culturing excised cotyledon explants with Agrobacterium tumefaciens. The best transformation efficiency was obtained on co-culture with the selective medium S2 (MS + 1.5 mg.L-1 6-BA + 0.2 mg.L-1 IAA + 500 mg.L-1 Cb) containing 20 mg. L-1 hygromycin. Roots were induced on S3 (1/2 MS + 20 mg.L-1 Hyg). Integration of FMD antigenic-determinant fused gene O21-O14 -A21-HBcAg into the genome of lettuce plants was confirmed by PCR and Southern blotting analysis. Expression of the fused gene at the transcriptional level was shown by RT-PCR.