转石蒜凝集素基因烟草的抗蚜虫性

利用农杆菌介导法将质粒pBILRA转化烟草(NicotianatabacumL.),该质粒含有由花椰菜花叶病毒35S启动子(CaMV35S)引导的筛选基因新霉素磷酸转移酶基因(nptII)及石蒜凝集素基因(lra)。通过卡那霉素筛选获得了25株独立转基因烟草植株。Westernblot分析表明,石蒜凝集素蛋白在不同转基因植株中表达量不同。对转基因T1代植株的遗传分析表明,lra基因在大多数独立转基因植株后代中以孟德尔31的分离比方式遗传。抗虫试验表明,表达较高水平石蒜凝集素蛋白的转基因烟草对桃蚜种群的生长具有明显的抑制作用。首次报道了表达石蒜凝集素基因的烟草对蚜虫具有抗性。石蒜凝集素基因可用于植物抗虫基因工程研究及应用。     

Cloning of a NaCl-induced fructose-1, 6-diphosphate aldolase cDNA from Dunaliella salina and its expression in tobacco

Itisaworld-widefocusthatsoilbecomesmoreandmoresaline.Aboutone-thirdirrigablelandbecomeslostallovertheworldforthereasonthatthesalinityistoohigh.Therefore,im-provingthesalt-toleranceofcropsseemsmoreandmoreimportant.Previousstudyonthemecha-nismofsaltresistantofplantshaspaidmoreattentiontohigherplants,whileonlyafewtoalgae.Studyintheseyearsshowedthatalgaehadthesimilarbiophysiologicalandbiochemicalreactiontoallhigherplantsinadversity[1].Soitisreliabletoapplyalgatorelatedstudy.Dunaliellasalina,akind…     

DNA芯片技术

ＤＮＡ芯片技术利用固定化在固体表面的高密度ＤＮＡ分子微点阵来进行生化分析。从生物体中提取的样品ＤＮＡ或ＲＮＡ作为靶分子,荧光或其它方法标记后,与微点阵上互补的探刈杂交,从而产生异源双链。因为在微点阵上,某一特定位置年的核苷酸序列是已知的,所以通过对微点阵每一位点的荧光强度进行检测,便可能样品进行定性及定量分析,检测技术包括激光共聚焦扫描和电耦合设备（ＣＣＤ）成像等等。ＤＮＡ芯片根据控针成分的不同大     

转石蒜凝集素基因烟草的抗蚜虫性

利用农杆菌介导法将质粒pBILRA转化烟草(Nicotianatabacum L.),该质粒含有由花椰菜花叶病毒35S启动子(CaMV35S)引导的筛选基因新霉素磷酸转移酶基因(nptⅡ)及石蒜凝集素基因(lra).通过卡那霉素筛选获得了25株独立转基因烟草植株.Western blot分析表明,石蒜凝集素蛋白在不同转基因植株中表达量不同.对转基因T1代植株的遗传分析表明,lra基因在大多数独立转基因植株后代中以孟德尔3:1的分离比方式遗传.抗虫试验表明,表达较高水平石蒜凝集素蛋白的转基因烟草对桃蚜种群的生长具有明显的抑制作用.首次报道了表达石蒜凝集素基因的烟草对蚜虫具有抗性.石蒜凝集素基因可用于植物抗虫基因工程研究及应用.     

受NaCl诱导的盐藻果糖-1,6-二磷酸醛缩酶cDNA的克隆及其在烟草中的表达

采用快速扩增cDNA末端(RACE)技术, 获得了盐藻(Dunaliella salina)受诱导表达的果糖-1,6-二磷酸醛缩酶全长cDNA (DsALDP). 蛋白质序列分析发现, DsALDP与许多植物叶绿体的果糖-1,6-二磷酸醛缩酶(AldP)具有较高的同源性(66%~73%). 系统进化分析进一步证明DsALDP与藻类的AldP亲缘关系最近. 就表达谱而言, DsALDP是NaCl诱导下新表达的转录本, 其表达水平随诱导时间的不同而呈显著变化. 筛选的DsALDP cDNA构建到双元载体pBI121并导入农杆菌用以转化烟草, 对4个T₁代转基因植株进行Southern检测, 证明DsALDP已整合入转基因植株的基因组. RT-PCR分析显示DsALDP在这些植株中均得到有效表达. 生物测试表明, T₁-1, T₁-2和T₁-3植株在100~200 mmol/L NaCl下仍保持醛缩酶活性, 且其脯氨酸含量均有不同程度的升高.     

Cloning of a NaCl-induced fructose-1, 6-diphosphate aldolase cDNA from<Emphasis Type="Italic">Dunaliella salina</Emphasis> and its expression in tobacco

Using Rapid Amplification of cDNA ends (RACE) technique, the full-length cDNA encoding a NaCl-induced fructose-1, 6-diphosphate aldolase (DsALDP) was obtained. It was shown that the DsALDP had a relatively high homology (66%–73%) to chloroplast fructose-1, 6-diphosphate aldolase (AldP) in many plants according to their amino acid sequences. The phylogenetic analysis further confirmed that AldP in alga is the nearest to DsALDP. As to its expression pattern,DsALDP was denovo synthesized by NaCl induction. Its expression level was significantly changed with inducing time. After the selectedDsALDP cDNA subcloned into a binary vector pBI121, the new construct was introduced into tobacco byAgrobacterium tumefaciens. The results of Southern blot and RT-PCR analysis of four transgenic T1 plants indicated thatDsALDP was integrated into genome of these transgenic plants and effectively expressed. Aldolase activities have been detected in T1-1, T1-2 and T1-3 plants by bioassay under 100–200 mmol/L NaCl. It was also observed that proline contents in them were differentially increased.