番茄LeNHX1基因植物过量表达载体的构建及其表达分析.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 20 10 2 LeNHX1 范晶 雷常安 黄明远 刘超 梁梓 陈丽萍 王永建 ( , 6 14004) : 从番茄幼苗中提取 RNA, 根据 NCB I 中番茄 L eNHX 1基因序 设计引物, 通过 RTPCR 获得了番茄 L eNHX 1基 因的 cDNA 序 , 包含 一个1 605 bp 的开放阅读框, 编码 534 个氨基酸将 cDNA 序 连接到植物过量表达载体 PBI121上, 对 所获得的重组质粒进行双酶切鉴定, 结果表明, 植物过量表达载体 PB I12 1LeNHX 1已构建成功半定量 RTPCR 结果表明 LeNHX 1基因在根茎和叶中均表达, 盐低温和脱落酸的诱导能提高 LeNHX 1基因的表达量, 推测番茄 LeNHX 1基因在逆境 应答中可能起着重要作用 : 番茄 L eNHX 1基因 植物过量表达载体 Expression Analysis and Construction of Plant Overexpression Vector ofTomato LeNHX1Gene Fan J ng Le Changan Huang M ngyuan L u Chao L ang Z Chen L p ng W ang Y ongj an (K ey Laboratory of P rotection and Utilization of B iodiversity, College of Chem istry and Lfi e Science, LeshanN ormal University, Leshan 614004) Abstrac:t T o tal RNA w as ex tracted from the tom a to young seed lng, cDNA w as obta ned by reve rse transc r pt on po lym erase cha n react on. P rm e rs w ere des gned acco rd ng to tom ato L eNHX 1 g ene sequence o f NCBI, then the target LeNHX 1 gene w as am pl f ed by RTPCR protoco.l the nucleot des fragm ent o f L eNHX 1 gene nc lud ng an OR F( 1605 bp) , encod ng 534 am no ac ds, The transcr pt of w h ch w as connec ted to p lant overexpress on vec to r PBI121. T he doub le d gest on o f recom b nant plasm d resu lt show ed that the plant overexpress on vectors w th L eNHX 1 w as constructed successfu lly. T he results of RTPCR show ed that L eNHX 1 w as expressed n roo ts, stem s and leaves, the express on level w as nduced by salt, low temperature and ABA stress. W e sugg est the L eNHX 1 gene could p lay an mportant ro le o f stress response n tom a to. K ey w ords: T oma to( Lycop ersicon esc lent m ) L eNHX 1 gene P lan t overexpress on vector + N a