Characterization of a PDR type ABC transporter gene from wheat （Triticum aestivum L.）

DON, as a virulence factor, plays an important role in the infection of Fusarium graminearum in wheat. The infection ability of F. graminearum depends on its capacity of producing DON. The production of DON by F. graminearum is significantly decreased in the wheat varieties with scab resistance. In this study, GeneChip analysis indicated that an EST encoding an ATP-binding cassette （ABC） transporter was up-regulated by 45 times in a wheat landrace Wangshuibai, which is resistant to DON accumulation. A pair of EST-derived primers were designed based on the EST sequence, and a clone was then isolated from a wheat genomic DNA TAC library. The TAC clone was sequenced using chromosome walking and gene prediction was conducted using Softberry. A cDNA clone of this gene was subsequently isolated from Wangshuibai induced by DON using gene-specific primers designed according to the untranslated sequence of the gene. The genome size of the gene is 7377 bp, consisting of 19 exons with coding sequences of 4308 bp. It encodes a protein with 1435 amino acid residues and the calculated molecular weight is about 161 kD. BLAST analysis indicated that the gene may belong to pleiotropic drug resistance （PDR） sub-family, and hence designated as TaPDR1 （Triticum aestivum pleiotropic drug resistance）. TaPDR1 was located on chromosome 5A of wheat using nullisomic-tetrasomic lines of Chinese Spring. TaPDR1 was up-regulated by induction of both DON and F. graminearum. Expression patterns of TaPDR1 were different in wild-type Wangshuibai and the fast-neutron induced Wangshuibai mutant lacking FHB1, a major QTL of FHB resistance and DON resistance in chromosome arm 3BS. These results suggested that TaPDR1 might be a candidate gene responsible for DON accumulation resistance. The expression profile showed that TaPDR1 expression was neither induced by hormones typically involved in biotic stress, such as JA and SA, nor by abiotic stresses, such as heat, cold, wounding and NaCI. However, TaPDR1 expression was regulated by Al^3＋ and [Ca^2＋], indicating that [Ca^2＋]1 might mediate the signal of TaPDR1 expression.     

稻瘟病菌中己糖载体蛋白的生物信息学分析

己糖载体蛋白是一种重要的糖载体蛋白,主要用于己糖的摄取和运输。本文通过生物信息学的方法,研究发现稻瘟病菌有67个可能的己糖载体蛋白。其中,MGG 06203、MGG 03620、MGG 15700、MGG 00040、MGG 13651、MGG 05946的氨基酸序列分别与Neurosporacrassa、Aspergillusnidulans和Colletrotrichumgramini-cola中已鉴定出的己糖载体蛋白高度同源。此外,稻瘟病菌不同的己糖载体蛋白基因在附着胞形成的各个阶段表达量存在很大的差异,尤其是其中有25个基因在稻瘟病菌侵染水稻48 h后有明显上调表达,表明它们可能参与了稻瘟病菌的致病过程。     

大鼠中性氨基酸转运蛋白SNAT1-HA融合蛋白的构建和表达(英文)

Na+偶联的中性氨基酸转运蛋白SNAT1是一种膜蛋白,属于转运蛋白第38家族(SLC38),在中枢神经系统(CNS)中起到重要的生理学作用.为了更容易地检测SNAT1在细胞膜上的表达,以pBK-CMVΔ-SNAT1为模板,用PCR,双酶切和T4连接酶连接的方法在SNAT1的C端加上了一个HA标签蛋白,构建了一个新质粒pBK-CMVΔ-SNAT1-HA.用该质粒瞬时转染人胚胎肾细胞(HEK293T),转染36 h以后,融合蛋白SNAT1-HA可以用HA抗体检测.Western blot结果显示在约54 kDa处有一条特异性条带,与预期分子量大小一致.利用重组质粒pBK-CMVΔ-SNAT1-HA可以更方便地检测到SNAT1在细胞膜上的表达与定位,为进一步研究SNAT1结构与功能奠定了基础.     

玉米秸秆横向输送装置结构运动参数试验研究

利用室内试验台,通过正交试验得到了影响输送综合评分的显著因素。通过单因素试验找出了各显著因素的较优参数值,在试验工况下,较优带速为1.484 7 m/s,较优齿端位为10 mm。对参数进行了的适应性试验,验证了较优参数组合的适应性。从而为玉米秸秆横向输送装置的设计提供参考依据。     

煤矿运输机的现状与发展

分析了SGZ-764／400型以下和SGZ-764／500型以上的小功率运输机在使用中存在的问题,阐明了SGZ-880／800等大功率运输机的优点及其发展前景。     

林可霉素转运蛋白基因lmrC的功能分析

为了研究腺苷三磷酸结合盒转运蛋白LmrC是否参与林可霉素的生物合成,通过同源重组在林可霉素高产菌株LC-G中构建了lmrC的缺失突变株XJJ7.高效液相色谱(HPLC)分析和抗性检测显示,相比出发菌株LC-G,XJJ7的林可霉素产量下降了55%,而且耐受林可霉素的最高质量浓度降低了50%.进一步实时荧光定量聚合酶链反应(PCR)分析发现,突变株XJJ7中林可霉素生物合成基因lmbA和lmbR的转录明显低于LC-G,但调控基因lmbU的转录不变.在LC-G中lmrC的过表达不能进一步提高产量,也不影响lmbA、lmbR和lmbU基因的转录,但过表达菌株对林可霉素的抗性显著增强.结果表明,lmrC可以通过影响生物合成基因的转录来调节林可霉素的产生,并赋予产生菌一定的抗性.