共查询到19条相似文献，搜索用时 59 毫秒
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为研究橡胶树中法尼基焦磷酸合酶基因(Hb FPS)的表达调控机制,采用酵母单杂交技术筛选了胶乳中与Hb FPS启动子结合的蛋白因子。将Hb FPS启动子与报告质粒p HIS2.1连接构建诱饵质粒p His-FPS,经筛选确定抑制背景表达的3-氨基三唑(3-aminotriazole,3-AT)浓度为10 mmol/L。同时以橡胶树胶乳m RNA为模板合成双链c DNA,并将双链c DNA、线性化的p GADT7Rec2载体和诱饵质粒p His-FPS共同转化酵母菌株Y187,两载体共转化效率为1.7×106 cfu/μg。筛选共得到阳性克隆36个,获得31条序列。对获得的c DNA序列进行生物信息学分析,得到2个转录因子序列,分别为类乙烯响应转录因子ERF003和Hb LMYC2。结果为进一步研究Hb FPS表达调控机制奠定基础。 相似文献
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以甘蓝型油菜热激蛋白p23（Brassica napus hsp23, Bnp23）为诱饵蛋白,用酵母双杂交LexA系统筛选拟南芥幼苗的AD-cDNA（activator domain-cDNA）文库,分离了39个酵母双杂交阳性克隆,其中两个阳性克隆的cDNA片段均编码拟南芥黑芥子酶结合蛋白（Arabidopsis thaliana PYK10 结合蛋白1,AtPBP1）。Blastp搜索到油菜黑芥子酶结合蛋白BnMBP6与拟南芥PBP1同源。经体外实验进一步证实AtPBP1和Bnp23之间存在相互作用。酵母双杂交实验也证实Bnp23能够与BnMBP6相互作用。 相似文献
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百脉根结瘤因子受体NFR5及其相互作用蛋白的研究 总被引：1，自引：0，他引：1
以豆科植物百脉根的mRNA为模板，利用RT-PCR方法，扩增到1.0kb结瘤因子受体5（nod factor receptor NFR5）蛋白激酶结构域的DNA片段（nfr5-pk），并将该片段克隆到酵母双杂交BD载体pGBKT7上。通过酵母双杂交技术，筛选接种根瘤菌的百脉根AD-cDNA酵母双杂交文库，从文库中分离到与nfr5-pk相互作用的221个阳性克隆。从这些阳性克隆酵母中分离AD质粒经反复验证，对26个确定阳性克隆的外源片段进行测序和同源性分析。通过NCBI数据库比对分析鉴定出小G蛋白Rop6等六个与NFR5-PK相互作用的不同蛋白，为进一步研究NFR5基因的功能和调控机制提供了材料。 相似文献
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水稻条纹病毒编码的Pc2蛋白在病毒侵染及宿主症状发生中起作用。研究其与寄主间的分子互作有助于揭示病毒侵染与致病的分子机制。分别将编码Pc2 N端（Pc2-N）与Pc2 C端（Pc2-C）的基因片段克隆到酵母双杂交诱饵载体pGBKT7上，并构建水稻叶片cDNA文库，然后分别用Pc2-N和Pc2-C为诱饵筛选文库寻找与其互作的寄主因子。结果表明，诱饵载体中插入的Pc2-N和Pc2-C编码框和氨基酸序列均正确。Pc2-N对酵母菌株AH109无毒性和自主激活能力，但Pc2-C对酵母具有细胞毒性。文库滴度为4×107 CFU/mL，且大多数插入片段在500~2 000 bp之间。利用Pc2-N为诱饵筛选水稻cDNA文库，获得有8个候选阳性克隆。经序列测定和BLAST分析结果表明，这些克隆共编码5种蛋白，主要为胁迫相关蛋白与光系统相关蛋白。 相似文献
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NPR1是水杨酸介导的系统获得性抗性（SAR）途径中不可或缺的一个转录辅助因子。在SA诱导下,NPR1被转运至细胞核内并与一些转录因子结合,以启动下游基因的表达。本研究通过PCR扩增获得拟南芥中NPR1基因CDS序列,经过限制性内切酶酶切后在T4连接酶作用下分别克隆至pGBKT7与pGADT7酵母表达载体上,并成功将其转化至Gold酵母菌株中。为利用酵母双杂交筛选NPR1参与形成的转录复合体中其它可能存在的蛋白质并阐明其在植物防御信号传导途径中的互作模式奠定了基础。 相似文献
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Cookie is one of the favorite cereal products which could be formulated by using various ingredients for enrichment. The aim of this study was to determine the effects of the inactivate yeast cells addition to cookie formulation in terms of glutathione (GSH) content, antioxidant activities, physicochemical and sensorial properties. Also, the cookies were produced with GSH (pure, 98%) addition to compare the effects of inactive yeast GSH with pure GSH on revealed properties. According to results, The GSH contents and the antioxidant activities increased in dough and cookie samples with inactive yeast addition compared to control and pure GSH added dough and cookie samples. The increases in GSH contents and antioxidant activities were observed after baking in all samples. The pure GSH addition increased the moisture content, spread ratio and L* value while decreased the hardness and b* values of the cookies. The contrary results were obtained by inactive yeast addition in terms of the physicochemical properties where the protein content of the cookies increased with inactive yeast addition by approximately 25%. The loss of GSH content decreased while the antioxidant activities and nutritional value increased in cookies by inactivate yeast addition. 相似文献
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玉米类Pto以及类Pti1基因参与多种逆境胁迫的信号传导。本研究分别将玉米ZmPto基因、ZmPti1a和ZmPti1b基因插入酵母表达载体pEG202和pJG4-5中,利用酵母双杂交的方法验证玉米类Pto以及类Pti1蛋白的相互作用。结果表明,无论是携带空载体还是携带一个(ZmPti1或ZmPto)融合蛋白的菌株都不能激活报告基因,只有同时携带ZmPti1和ZmPto蛋白的菌株可以激活报告基因,初步确定ZmPto蛋白与ZmPti1蛋白存在相互作用。 相似文献
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为了研究植物核蛋白的组成及其在各种环境条件下的动态变化,利用核蛋白的核定位信号(nuclear localization signals,NLS)筛选编码核蛋白的基因,建立了一套快速、省力、低成本的核蛋白筛选系统(nuclear transportation trap,NTT).通过鉴定发现,将合成的SV40蛋白大T抗原的核定位信号序列插入筛选载体的多克隆位点,转化酵母EGY48,插入核定位信号的筛选载体转化酵母后可以在选择性培养基(SD/His-/Leu-)上生长,而没有插入核定位信号的筛选载体转化酵母后不能在选择性培养基上生长,从而证明此核蛋白筛选系统有效.比较实验表明,本研究构建的核蛋白筛选系统与已报道的系统相比筛选效率更高.利用此系统从cDNA文库中分离编码核蛋白的基因,对于研究核蛋白的动态组成、了解核蛋白基因在调控细胞发育及其环境胁迫条件下的作用机制具有重要的意义. 相似文献
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Rheofermentometer parameters and bread specific volume of frozen sweet dough influenced by ingredients and dough mixing temperature 总被引：2，自引：0，他引：2
Weining Huang Yangsoo Kim Xianyu Li Patricia Rayas-Duarte 《Journal of Cereal Science》2008,48(3):639-646
Response surface methodology described the effects of salt, lactic acid, shortening, and exogenous trehalose and dough mixing temperature (DMT) and their interactions on the three rheological and fermentation parameters. These included maximum dough height (Hm), maximum height of gas release (Hm′) and CO2 production, measured by the Rheofermentometer F3, and bread specific volume (Sp. Vol.) of frozen sweet dough. The models could estimate the four parameters with R2 values of 0.76, 0.69, 0.93, and 0.59, respectively. Salt significantly influenced all four parameters in a negative way. DMT affected positively the Hm and Sp. Vol. of bread. Lactic acid affected Hm only, but its interactions with other variables influenced all four parameters. Shortening level affected Hm′ and CO2 production positively and Sp. Vol. negatively. The added exogenous trehalose improved Hm, Hm′, and CO2 production significantly, but not the Sp. Vol. of bread. Among the three Rheofermentometer parameters, Hm showed the highest correlation with Sp. Vol. (R2 = 0.75). DMT for the maximum Hm and Sp. Vol. varied with the level of other ingredients. Trehalose alone could not overcome the challenges in a sweet frozen dough system to improve the Sp. Vol., and its combined effects with other ingredients will need to be evaluated to restore the impaired gas retention of the frozen sweet dough. 相似文献
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When used in bread dough systems, glucose oxidase (GO) and pyranose oxidase (P2O) generate H2O2 from O2. We here studied their potential to improve dough and bread characteristics. Neither GO nor P2O significantly affected the volume of straight dough bread produced with fermentation and proofing times of respectively 90 and 36 min at dosages up to 0.50 nkat/g flour. Supplementation with 1.00 nkat/g flour of GO or P2O significantly decreased bread loaf volume. The resistance of dough (fermented for 20 min and proofed for 56 min) to an applied shock was substantially improved by inclusion of 0.08, 0.25, 0.50 or 1.00 nkat/g flour of GO or P2O in the dough recipe. Thus, the proofed doughs showed significantly less collapse and the resultant breads had higher loaf volumes than did the reference breads. Yeast probably exerts an oxidizing effect on dough, which, depending on the exact breadmaking protocol used, might veil the positive oxidizing effect of the enzymes on dough properties during prolonged fermentation. 相似文献
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Freezing deteriorates the baking quality of frozen bread dough by causing lethal injury to yeast cells and depolymerization to the gluten network. To investigate the potential of biogenic ice nucleators in frozen food applications, the effect of extracellular ice nucleators (ECINs) from Erwinia herbicola on the baking quality of frozen dough upon three freeze/thaw cycles were investigated. With addition of ECINs to the activity of 2.4 × 106 units per gram of dough, hardening of bread crumb caused by three freeze/thaw cycles was alleviated by about 50% compared to the control. Additionally, the bread from frozen dough with added ECINs showed 50% larger specific volume compared to the control. The mechanism of cryoprotective effects from ECINs was possibly that ECINs helped in preserving the viability of yeast cells during freeze/thaw cycles. ECINs were able to improve the viability of log-phase and stationary-phase yeast cells in suspensions by about 100 and 10 fold, respectively, and viability of yeast in the frozen dough by 17%. This study revealed the potential of ECINs as a cryoprotectant for applications in the food and biotechnology industries. 相似文献