香蕉ACC氧化酶基因(MAO3)的克隆及其表达特性分析   总被引：3，自引：0，他引：3  

黄俊生  王华  张世清 《园艺学报》2005,32(5):807-811

 根据同源扩增得到香蕉(Musa acuminata) ACC氧化酶基因(MAO3) 的核心部分, 再通过3'和5'RACE扩增上、下游序列以及Genome-Walker的方法得到启动子部分, 共获得3 718 bp长度的序列。将所得结果进行聚类分析, 发现香蕉中ACC氧化酶的氨基酸序列非常保守, 各序列间的同一性高达99% ,与单子叶植物和双子叶植物中ACC氧化酶的氨基酸序列的同源性在66.7%～71.8%之间。组织原位杂交试验表明MAO3基因的表达具有组织特异性, 初步认为是在韧皮部筛管组织中特异表达。运用实时荧光定量PCR技术, 实时监测到了MAO3基因和香蕉乙烯受体基因ERS2受机械伤诱导的定量变化。  相似文献   

鸡传染性腔上囊病病毒VP2基因在昆虫细胞中的表达   总被引：4，自引：0，他引：4  

李迎晓秦爱建  刘红梅邵红霞金文杰  刘岳龙 《中国兽医科技》2005,35(12):933-936

为了深入研究鸡传染性腔上囊病病毒（IBDV）VP2基因的结构和功能,利用Bac-to-Bac系统研制出含有VP2基因的重组杆状病毒rBac-VP2,将rBac-VP2感染Sf9细胞,并用抗IBDV VP2特异性单克隆抗体经间接免疫荧光试验检测,证实感染重组病毒Sf9细胞能高效表达IBDV VP2基因产物;Western-blotting分析结果表明,VP2基因表达产物的分子质量约为40 ku.  相似文献   

Quick detection of sex determining region protein gene in mouse fetuses     

《园艺学报》2003,19(5):622-626

AIM: To detect quickly the Y-chromosome specific sex determining region protein (Sry) gene in mouse fetuses on embryonic day 14.5 with a PCR method. METHODS: We designed specific primers with the OLIGO 5. 0 software. Templates were prepared in 30 minutes by the following way. About 1 mg embryonic tissue but not fetal liver was suspended, and treated with 200μL of lysis buffer, consisting of PCR buffer containing 20 mg/L proteinase K, 0. 5% NP-40, and 0.05% Tween 40, at 60°C for 15 minutes, heated for 5 minutes at 100 °C, 10μL was used as template. The PCR react ion was performed in 50μL, using two sets of primers specific for Sry gene (chromosome Y) and IL-3 gene (chromosome 11) . PCR conditions and cycle numbers were optimized. The assessment of the results was done by electrophoresis in 3% agarose run at high voltage. The specificity of the method was conf irmed by fluorescent in situ hybridization (FISH) using a specific male probe on embryonic tissue cells. RESULTS: Electrophoresis showed that PCR product of male control DNA consisted of a 649 bp product representing the IL-3 gene and a 444 bp product representing the Y-specific Sry gene, female control DNA only one 649 bp product. Fetuses with two bands matching those as seen inmale control DNA are the presumpt ive male fetuses. Fetuses, only the IL-3-associated 649 bp band, are the presumptive female fetuses. These were confirmed by FISH. The ent ire procedure took <3. 5 h. CONCLUSION: The established PCR assay offers a quick, simple, accurate, and sensitive detection of sex determining region protein gene in mouse fetuses. This method allowed the preparation and culture of pure male and female hematopoietic stem cells from fetal tissue.  相似文献   

Amino Acid Changes in Pepper mild mottle virus Coat Protein That Affect L 3 Gene-mediated Resistance in Pepper     

Hiroyuki HAMADA  Shigeharu TAKEUCHI  Akinori KIBA  Shinya TSUDA  Yasufumi HIKICHI  Tetsuro OKUNO 《Journal of General Plant Pathology》2002,68(2):155-162

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Gene Flow Analysis of Phytophthora porri Reveals a New Species: Phytophthora brassicae Sp. Nov.     

Willem A. Man in 't Veld  Arthur W.A.M. de Cock  Elena Ilieva  C. André Lévesque 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(1):51-62

Isozyme analysis and sequence analysis of the internal transcribed spacer regions (ITS-1 and ITS-2) and the 5.8S subunit of the ribosomal DNA gene repeat were used to examine whether isolates of Phytophthora porri from Allium and Brassica represent a single homogeneous species. Twenty-six strains of P. porri, 16 strains isolated from the genus Allium, and 10 strains isolated from the genus Brassica, were analyzed using malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH) and lactate dehydrogenase (LDH), represented altogether by four putative loci (Mdh-2, Idh-1, Idh-2, and Ldh-2). Isozyme analysis revealed that strains isolated from Allium contained five private alleles at three isozyme loci (Ldh-2 ⁸³, Ldh-2 ¹⁰⁴, Idh-1 ¹⁰⁸, Idh-1 ¹¹², and Idh-2 ⁹⁸), whereas six different alleles were observed at four isozyme loci (Ldh-2 ⁸⁵, Ldh-2 ¹⁰⁰, Ldh-2 ¹¹⁴, Idh-1 ¹⁰⁰, Idh-2 ¹⁰⁰, and Mdh-2 ¹¹¹) in strains obtained from Brassica. The heterozygosity at the Ldh-2 locus, differing in allele composition, however, between strains from Allium and Brassica, was present in all strains, indicating that it is probably fixed. Sequence analysis of the ITS regions and the 5.8S subunit showed consistent differences between isolates from Allium and isolates from Brassica. Based on isozyme data, ITS sequence analysis and formerly published differences in restriction enzyme patterns of mitochondrial DNA, morphology and pathogenicity, it was concluded that the isolates of P. porri Foister did not represent a homogeneous species. Isolates from Brassica constitute a distinct species which is described here as P. brassicae sp. nov. It was inferred from isozyme patterns, which were in no case intermediate between the two species, that P. porri and P. brassicae do not hybridize and are reproductively isolated by barriers to gene flow.  相似文献   

Bt对小地考虑呼吸作用的影响   总被引：5，自引：0，他引：5  

李芳  谌建春等 《昆虫天敌》2002,24(1):15-19

利用红外CO2分析仪测定小地老虎幼虫呼吸节律的变化结果显示：小地老虎幼虫呼吸节律属于不规则波动型，取食含Bt烟叶的幼虫呼出的CO2量显著低于对照组，且呼吸节律的波动性也明显增大，说明小地考虑幼虫取食Bt后，呼吸代谢受到一定干扰，出现呼吸水平降低及代谢紊乱现象。  相似文献   

新疆棉区转Bt基因棉对棉田主要害虫及其天敌种群数量的影响   总被引：19，自引：0，他引：19  

孙长贵  徐静  张青文  封红兵  汪飞  宋荣 《中国生物防治》2002,18(3):106-110

20 0 0～ 2 0 0 1年 ,在新疆棉区系统地研究了转Bt基因棉田主要害虫及其天敌的发生规律。结果表明 :转Bt基因棉对棉铃虫有良好的控制作用 ,发生高峰期百株落卵量和百株幼虫数量都显著低于常规对照 ,转Bt自控田和转Bt化防田棉蚜和棉盲蝽发生数量则显著高于常规棉对照田。在天敌方面 ,转Bt自控田和转Bt化防田较常规棉对照田 ,丽草蛉种群数量有所增加 ,而小花蝽和黑食蚜盲蝽等 5种捕食性和几种寄生性天敌数量都有所下降。转Bt基因棉田棉蚜和棉盲蝽等刺吸性害虫发生数量加重而多种天敌数量减少的趋势值得引起重视。鉴于新疆地区棉铃虫发生数量较低目前无需专门防治的特点 ,作者建议 :可以考虑推广农艺性状较好的常规棉品种并保护天敌 ,以达到控制害虫、丰产丰收的目的。  相似文献   

苏云金芽孢杆菌cry基因在大肠杆菌中表达产物和生物活性   总被引：4，自引：0，他引：4  

韩岚岚  宋福平  李长友  张杰  赵奎军  黄大昉 《植物保护》2002,28(5):5-9

研究了几种Bt cry基因于大肠杆菌(Escherichia coli)中表达产物在pH 10.0的50mmol/L碳酸钠和20mmol/L乙醇胺溶解液中的溶解性 ,发现同样的Cry蛋白在碳酸钠中的溶解度大于乙醇胺。通过胰蛋白酶消化 ,明确Cry1Ca7、Cry1Ia8酶解产物为 38kD多肽 ;Cry1Ie1、Cry1Cb2、Cry2Ab4酶解产物为 41kD多肽 ;Cry1Ac酶解产物为60kD多肽。采用FPLC层析方法对 6种原毒素及其酶解后得到的毒素多肽进行了分离纯化 ,比较了原毒素和毒素的杀虫活性的差异。其结果表明 ,Cry1Ac的原毒素和毒素对棉铃虫初孵幼虫的校正死亡率均为 100% ,Cry2Ab4的原毒素的毒力高于其酶解毒素。  相似文献   

一种监测棉铃虫对Bt杀虫晶体蛋白抗性的技术   总被引：1，自引：0，他引：1  

范贤林  芮昌辉  孟香清  梁桂梅  李永平 《植物保护学报》2002,29(3):254-258

研究了以诊断试剂盒监测棉铃虫对苏云金杆菌(Bt)杀虫晶体蛋白(ICP)抗性的技术。将1日龄幼虫在含诊断剂量的饲料中(每毫升饲料含Bt ICP0.01mg)取食7天,以生长至3龄以上的个体作为抗性个体的判断指标;筛选出人工饲料中防腐剂剂量的最佳值为每350毫升饲料中含丙酸2.5ml。经对多个棉铃虫不同抗性种群的验证试验,证明诊断试剂盒检测抗性个体频率较为精确且应用简便。  相似文献   

转 Bt基因抗虫棉对棉大卷叶螟抗性的研究   总被引：7，自引：0，他引：7  

王厚振  肖云丽  郑成民  王书友  孟妍遐 《中国植保导刊》2002,22(3):21-22

:转Bt基因抗虫棉对棉铃虫有很好的抗性,对棉大卷叶螟(SyleptaderogataFabricius)的抗性未见报道。作者研究表明,棉大卷叶螟在棉田为聚集分布型,低龄幼虫有集中为害习性;抗虫棉对其有很好的抗性,平均虫株率为2.2%,较常规棉中棉所12降低96.6%;百株虫量为133.8头,较常规棉降低88.4%。  相似文献