猪SENP1基因克隆、生物信息学分析与表达研究

试验旨在对猪SUMO特异性蛋白酶1(sentrin-specific protease 1,SENP1)基因CDS区进行克隆和生物信息学分析,并探讨SENP1基因在乙脑病毒(Japanese encephalitis virus,JEV)感染PK15细胞后的表达情况。根据GenBank中公布的猪SENP1基因预测序列(登录号:XM_013997974.2)设计特异性引物,通过RT-PCR和T/A克隆技术对猪SENP1基因CDS区序列进行克隆测序,并进行生物信息学分析,利用实时荧光定量PCR分析猪SENP1基因在JEV感染PK15细胞不同时间点(0、12、24、36、48 h)的表达情况。结果显示,猪SENP1基因CDS序列全长2 034 bp,共编码677个氨基酸。序列比对和系统进化树分析结果表明,猪SENP1蛋白序列和山羊、犬、人、牛的相似性分别为94.8%、94.2%、93.9%和93.8%,说明在这些物种中SENP1的保守性较强;猪与犬的SENP1分子亲缘关系较近。生物信息学分析结果显示,猪SENP1蛋白相对分子质量为76 825.76,等电点为8.53,体外半衰期为30 h,不稳定系数为53.59,总平均亲水指数为－0.622。SENP1蛋白不含信号肽序列,无跨膜结构域。二级结构分析结果显示,SENP1蛋白中α-螺旋、无规则卷曲、延伸链和β-转角分别占31.31%、46.68%、16.25%和5.76%。三级结构分析结果显示,猪SENP1蛋白中存在8个α-螺旋区和7个β-转角区。实时荧光定量PCR结果显示,猪SENP1基因在JEV感染的PK15细胞中呈现上调表达趋势,其中在感染后48 h SENP1基因表达量显著高于对照组(P<0.05)。本试验结果为后续深入研究SENP1基因功能提供了参考。     

Energy metabolism and apoptotic effect of microwave radiation on rat myocardial cells

AIM: To investigate the effect of microwave radiation at different intensities on the rat myocardium and its possible mechanism.METHODS: The rats were radiated by the intensity of 500, 1 000, 1 500 and 2 000 W/m² with 2 450 MHz microwave for 6 min. The heart tissue was collected 6 h after microwave radiation. ATP and mitochondria complex Ⅳ and Ⅴ were measured. The changes of the tissue structures were observed under transmission electron microscope. The apoptosis of the myocardial cells was detected by a cell analyzer. The protein level of cleaved caspase-3 was determined by Western blotting.RESULTS: The concentration of ATP and activity of mitochondria complex Ⅳand Ⅴ signi-ficantly decreased compared with control group in the cardiac tissues. The decreased number, morphological abnormalities such as dissolved cavitation, matrix and obvious tumefaction of mitochondria were observed under transmission electron microscope. The microwave radiation induced the apoptosis of myocardial cells in the rats. The cell apoptotic rate and the protein level of cleaved caspase-3 increased with increasing intensity of microwave radiation(P<0.05).CONCLUSION: Microwave radiation has obvious injury effect on the rat heart, which can cause cardiac energy metabolism dysregulation and cardiac myocyte apoptosis.     

南瓜新品种科栗1号的选育

科栗1号是以S31-1为母本、S4-1为父本配制而成的南瓜一代杂种。植株蔓生,长势中等;分枝较少,叶小,节间短;第1雌花节位第9节左右;嫩瓜扁圆形,瓜皮绿色覆绿色斑点和浅色纵条斑纹,老瓜皮墨绿色。嫩瓜单瓜质量0.5~1.5kg,果实横径9cm左右,纵径13cm左右,糯性好,粉质,品质佳;春季露地和秋季设施栽培嫩瓜产量1500~2000kg·(667m2)-1,早春设施栽培嫩瓜产量3000kg·(667m2)-1左右。适宜浙江省及栽培条件相似的地区推广种植。     

Expression,Purification and Activity Evaluation of Mb1230 Gene of Mycobacterium bovis

To explore the value of Mb1230 protein of Mycobacterium bovis in the diagnosis of bovine tuberculosis,we obtained the Mb1230 gene by PCR and constructed recombinant plasmid pET-22b-Mb1230.Recombinant Mb1230 protein was obtained by IPTG induction and purified by affinity chromatography.The activity of the recombinant protein was evaluated by TST test,IGRA test and indirect ELISA.The size of the recombinant protein matched with the theoretical value proved by SDS-PAGE;Western blotting result showed that the recombinant protein could react with mouse anti-His antibody,and had specific band;The results of TST test,IGRA test and indirect ELISA test also showed the recombinant protein had antigenic activity.The results indicated the recombinant protein Mb1230 had good B cell and T cell activity,so,it had the potential application in the diagnosis of bovine tuberculosis.     

中国石油海外管道完整性管理的必要性和挑战

针对国家油气战略通道安全平稳运行的需要,基于中国石油海外油气管道的基本特点,分析了海外油气管道实施完整性管理的必要性。根据中国石油海外管道安全运行的技术需求和油气管道完整性管理技术的特征,分析了中国石油海外油气管道实施完整性管理面临的技术挑战。为提高海外管道安全水平,避免海外投资遭受损失,需尽快开展海外管道的完整性技术应用工作。在具体实施过程中,需要与国内管道完整性管理技术研究及应用单位相结合,充分发挥中国石油集团公司整体技术优势。     

柞蚕新品种永青的选育

柞蚕新品种永青是以“青六号”为基础材料,利用系统选育手段,采用分区饲养、系统分离、定向培育的方法,同时兼顾各项经济指标的综合平衡,通过9年18代的系统选育,育成的具有青黄蚕血统的柞蚕新品种。选育结果：全茧量12.47g,千粒茧重11.99kg,虫蛹统一生命率94.81%;与吉柞88-2、方山黄杂交,增产幅度达15.5%以上。     

覆膜滴灌玉米产量和农艺性状相关性分析

为明确在覆膜滴灌条件下玉米品系(种)主要农艺性状与产量的关系,应用DPS软件对15个玉米杂交品系(种)的主要农艺性状和产量性状进行相关性、灰色关联度等分析,结果表明:籽粒重、穗重、产量的变异系数较大;玉米产量与生育期、籽粒水分含量、穗重、籽粒重呈显著正相关关系;聚类分析可将参试的15个品系(种)划分为3类,第Ⅲ类表现良好,值得进行推广种植。灰色关联度分析表明,玉米籽粒重、穗重、生育期、籽粒水分含量等性状是影响玉米产量的主导因素,在玉米高产育种中,应以其为主攻目标,提高玉米后代的有效选择,加速高产育种进程。     

甘蔗引进新品种(系)的试验表现及评价

为筛选出高产、高糖、综合性状表现较好的甘蔗新品种充实蔗区,对17个国内外甘蔗引进新品种(系)进行了1年新植1年宿根种性比较试验。结果表明,平均蔗茎产量显著高于新台糖22号的参试品种有SP80-3280、K86-110、ISD24、FR93-344、CP84-1591、VMC88-354和K82-129,增产10.54%~21.47%。平均蔗糖分含量高于新台糖22号的参试品种有VMC71-39、CP94-1100、FR93-435和云蔗03-258,蔗糖分含量提高0.2~1.4个百分点。     

北京水系中腐霉和疫腐霉种类调查

为研究北京地区水系中腐霉和疫腐霉种类分布,采用叶饵法从北京水系中诱集菌株,共分离到腐霉、疫腐霉182株,利用Single-strand conformation polymorphism (SSCP)进行分析,筛选出不同基因型的代表菌株,并依据形态学特征及ITS序列对这些菌株进行种类鉴定。结果共发现6个已知种和3个可能的新种,其中:湖滨疫腐霉Phytopythium litorale和果胶裂解腐霉Pythium pectinolyticum为国内新记录种;旋柄疫腐霉Pp.helicoides、异丝腐霉P.diclinum、簇囊腐霉P.torulosum和袋囊腐霉P.marsipium为北京地区新记录种。研究结果可为未来监测北京地区水系中病原菌的分布情况提供基础数据。