玉米大斑病菌StSNF1的基因组定位、蛋白质结构预测及表达分析

为了明确StSNF1在玉米大斑病菌基因组中的位置,解析该基因编码蛋白的结构特征,探究该基因在侵染寄主的不同时期、分生孢子萌发侵染过程中以及不同碳源培养下的表达情况。结果表明,该基因ID号为008026214,全长3 046 bp,位于scaffold_17负链的97 793-100 838位置。StSNF1与玉米圆斑病菌SNF1同源关系较近,由877个氨基酸残基编码而成。StSNF1蛋白具有氮端的蛋白激酶结构域、氮端的碳代谢产物去阻遏蛋白激酶结构域和碳端的激酶相关结构域。在蛋白激酶结构域内,具有ATP结合位点和丝氨酸/苏氨酸活性位点。Real-time PCR结果表明,StSNF1在侵染后期高表达,72 h表达量最高;StSNF1在分生孢子萌发24 h时(即侵入丝形成时期)表达量最高;StSNF1在蔗糖为单一碳源的培养基中表达量最高,果胶培养基次之。综上所述,StSNF1与侵染寄主和碳源利用密切相关,在侵染后期发挥作用,利用寄主细胞内的非发酵型碳源进行次级侵染。     

Role of cytokeratin 8 on changes of intestinal epithelial permeability induced by corticotropin-releasing factor

AIM: To investigate the role of cytokeratin 8 (CK8) on the change of intercellular permeability of intestinal epithelial cells induced by corticotropin-releasing factor (CRF). METHODS: The expression levels of CRF receptor 1 (CRFR1) and CRFR2 on human colon adenocarcinoma HT29 cell surface were determined by immunofluorescence staining. After treatment with 100 nmol/L CRF for 72 h, the translocation of FITC-labelled dextran was measured in a Transwell chamber. The structural changes of tight junctions were observed under transmission electron microscope. The expression levels of CK8, and tight junction proteins ZO-1 and occludin were determined by Western blot. The activity of protein kinase C (PKC) was detected by ELISA. Furthermore, the effects of CRF on intestinal epithelial permeability were examined in CK8-silencing HT29 cells, which were constructed by infection with sh-CK8 lentivirus. RESULTS: CRF treatment increased the permeability of FITC-labelled dextran (P<0.05), caused the opening of tight junctions, and induced increased fluorescence intensity of CK8. The expression levels of occludin and ZO-1 were down-regulated (P<0.05). PKC activity was decreased at 1 h after CRF treatment (P<0.05). CRF-induced increase in the permeability and down-regulation of occludin were not blocked by CK8 silencing. Nevertheless,CK8 silencing blocked the effects of CRF regarding the decrease in the expression levels of ZO-1 and the increase in PKC activity (P<0.05). CONCLUSION: CK8 may be involved in CRF-induced increase in intestinal epithelial permeability by inhibiting the activity of PKC, and there may be other signaling pathways involved.     

FAM3C activates Akt to promote viability of oral squamous-cell carcinoma cells

AIM: To investigate the expression and roles of family with sequence similarity 3, member C (FAM3C) in oral squamous-cell carcinoma cells. METHODS: The mRNA and protein expression levels of FAM3C in dysplastic oral keratinocyte (DOK) and oral squamous-cell carcinoma WSU-HN6 cells were detected by RT-qPCR and Western blot. The WSU-HN6 cells were treated with siFAM3C or FAM3C antibody. After 24, 48 and 72 h, the viability of WSU-HN6 cells was measured by CCK-8 assay, and the activation of protein kinase B (Akt) was detected by Western blot. Adenovirus was used to mediate over-expression of FAM3C in the DOK cells. The DOK cell viability was measured by CCK-8 assay after adenovirus infection for 24, 48 and 72 h, and the activation of Akt was detected by Western blot. RESULTS: Compared with the DOK cells, the mRNA and protein levels of FAM3C were significantly increased in the WSU-HN6 cells (P<0.05). The viability of WSU-HN6 cells transfected with siFAM3C was significantly inhibited at 48 h and 72 h (P<0.05). siFAM3C treatment inhibited the activation of Akt (P<0.05). FAM3C antibody treatment also suppressed the viability of the WSU-HN6 cells at 48 h and 72 h and the activation of Akt (P<0.05). Over-expression of FAM3C in the DOK cells promoted the cell viability at 48 h and 72 h and activated Akt (P<0.05). CONCLUSION: FAM3C might promote oral squamous-cell carcinoma cell growth by activating Akt.     

犊牛代用乳中蛋白质原料应用研究进展

代用乳是犊牛主要的液体饲料来源之一,与全乳和乳副产品相比不仅降低了犊牛生产成本,而且更适合犊牛饲养设备的操作和使用.在以代用乳为主要饲料来源培育犊牛过程中,代用乳中蛋白质的营养作用是决定饲养效果和经济性的关键性因素.本文对代用乳常用蛋白质的原料特性等方面做了简要综述.     

花生乳状液体系中蛋白质的酶解动力学研究

采用碱性蛋白酶Alcalase 2.4L水解花生乳状液体系中蛋白质,研究了蛋白酶在乳状液体系中的水解特性。结果表明：蛋白质水解度随着初始酶浓度的增加而提高,在酶浓度一定的情况下水解度随着初始底物浓度的提高而降低。蛋白质水解度与乳状液的破乳率存在显著正相关(r=0.983)。在花生乳状液体系中Alcalase 2.4L的水解动力学参数Km=0.0698mol/L,Vmax=3.71×10-4mol/(min·L)。由实验数据推导出蛋白质酶解初级阶段的动力学方程,确定蛋白质水解和乳状液破乳的临界初始底物质量浓度为8.73g/L（加酶量为0.05%）。在酶和底物初始浓度以及反应时间确定的前提下,通过动力学方程可预测破乳过程中蛋白质的水解度及破乳率。     

酒糟菌体蛋白饲料加工技术的应用研究

微生物技术开发蛋白饲料是一项高科技项目，是国家要求重点推广的项目。以山西杏花村汾酒（集团）公司为生产基地，利用该公司生产白酒的下脚料——酒糟为原料，提出了切实可行的工厂化菌体蛋白饲料生产工艺，并进行了成套设备的研制。该项技术的应用实现了酒糟的高效再利用，为解决我国饲料行业蛋白源缺乏提供了一条新途径。     

一种土壤微生物总DNA的高效提取方法

获得高浓度、大片段、多样性程度高的土壤微生物总DNA 是研究土壤微生物群落结构的分子生态学基础。本文采用间接法(菌体细胞回收法)提取红壤地区两种土壤类型的土壤微生物总DNA,定量计算其回收率,并与直接法(细胞原位裂解法)比较了提取效率和纯度。结果表明:红壤地区2种土壤每克干土的总DNA提取量,间接法约为0.34和0.53礸/g干土,直接法约为13.62和24.32礸/g干土;间接法的提取效率低于直接法,但所得DNA片段较大,且Sau 3AⅠ 酶切和16 S rDNA通用引物PCR扩增结果显示,间接法比直接法更能有效地去除土壤中的某些抑制剂,所得总DNA的纯度更高,有利于后续操作。     

杠柳根皮粗提液对菜青虫生物活性的影响

试验研究杠柳根皮粗提液对菜青虫生物活性的影响测定结果表明,杠柳根皮乙醇提取液对菜青虫的生物活性影响高于杠柳根皮氯仿提取液和石油醚提取液,杠柳根皮乙醇提取液对菜青虫幼虫有较强的拒食作用和生长抑制作用,胃毒作用一般。用100倍液杠柳根皮乙醇提取液浸叶饲喂3龄、4龄、5龄菜青虫幼虫后24h其拒食率分别为94.9%、87.8%和92.7%,拒食后幼虫的化蛹率和蛹重均比对照显著降低。100倍液杠柳根皮乙醇提取液浸叶饲喂4龄菜青虫幼虫后24h和72h其生长抑制率分别为96.6%和82.5%。杠柳根皮乙醇提取液对菜青虫成虫产卵具有明显忌避作用,用50~100倍液杠柳根皮乙醇提取液喷雾处理油菜苗后2~4d其产卵忌避率达89.5%以上。     

绵羊对4种蛋白饲料日粮营养物质消化动态变化的比较

试验采用3头35 kg左右的美利奴绵羊研究不同蛋白饲料日粮对绵羊营养物质消化的影响。为自身对照试验设计,整个试验分4期进行,每期3头羊饲喂同一种蛋白饲料日粮。4种蛋白来源分别为棉籽粕、大豆粕、葵粕和菜籽粕。试验结果表明:绵羊采食蛋白来源为棉籽粕的日粮时,干物质、有机物和氮采食量最高,而菜籽粕采食量最低。饲喂棉籽粕日粮,干物质、有机物和纤维素十二指肠流量显著高于大豆粕日粮和菜籽粕日粮(P<0.05)。大豆粕日粮粗蛋白在前胃的消失率最高为33%,其余3组日粮粗蛋白前胃的降解率为16%~20%。到达小肠的菌体蛋白(MCP)量各组间无差异,而到达小肠过瘤胃饲料蛋白的量(率)棉籽粕组显著高于葵粕组、菜籽粕组、大豆粕组。大豆粕组粗蛋白在小肠的表观消化率最低为48%,其余3组日粮粗蛋白小肠表观降解率均为66%。     

杂交水稻品种指纹鉴定研究进展

从蛋白质指纹和DNA指纹两个方面,综述了杂交水稻品种指纹鉴定研究的进展和不足。对SSR标记在品种指纹鉴定上的应用前景,单一特异标记的筛选和实验室标准检测技术的建立作了探讨和展望,以利于加快杂交水稻品种指纹鉴定的发展和应用。