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.     

南瓜银叶突变体48a的表型特征及其遗传学分析

南瓜银叶突变体48a是在嫩食型中国南瓜中分离筛选到的稳定遗传自交系,银色叶不仅可以作为标记性状应用到育种中,还可为南瓜抗虫、抗病、耐寒等一系列研究提供重要的材料基础。笔者对银叶突变体的表型特征及叶片的解剖结构进行鉴定分析,发现植株整体长势、熟性与野生型无明显差异;成熟叶片正面全部呈银灰色,叶绿素含量明显降低,叶片上表皮细胞与栅栏细胞间明显剥离,存在明显的空隙。利用突变体48a和野生型49a南瓜自交系构建的六世代遗传群体,调查发现F2的绿叶与银叶符合3∶1的分离比,回交群体BC1P1分离比符合1∶1,表明南瓜银色叶性状由单隐性基因控制。     

Lipid-rich bovine serum albumin improves the viability and hatching ability of porcine blastocysts produced in vitro

The effects of lipid-rich bovine serum albumin (LR-BSA) on the development of porcine blastocysts produced in vitro were examined. Addition of 0.5 to 5 mg/ml LR-BSA to porcine blastocyst medium (PBM) from Day 5 (Day 0 = in vitro fertilization) significantly increased the hatching rates of blastocysts on Day 7 and the total cell numbers in Day-7 blastocysts. When Day-5 blastocysts were cultured with PBM alone, PBM containing LR-BSA, recombinant human serum albumin or fatty acid-free BSA, addition of LR-BSA significantly enhanced hatching rates and the cell number in blastocysts that survived compared with other treatments. The diameter, ATP content and numbers of both inner cell mass and total cells in Day-6 and Day-7 blastocysts cultured with PBM containing LR-BSA were significantly higher than in blastocysts cultured with PBM alone, whereas LR-BSA had no effect on mitochondrial membrane potential. The mRNA levels of enzymes involved in fatty acid metabolism and β-oxidation (ACSL1, ACSL3, CPT1, CPT2 and KAT) in Day-7 blastocysts were significantly upregulated by the addition of LR-BSA. The results indicated that LR-BSA enhanced hatching ability and quality of porcine blastocysts produced in vitro, as determined by ATP content, blastocyst diameter and expression levels of the specific genes, suggesting that the stimulatory effects of LR-BSA arise from lipids bound to albumin.     

辣椒疫霉漆酶基因Pclac3克隆表达

辣椒疫霉是一种侵染性植物病原,能引起多种茄科及葫芦科植物的病害。利用同源克隆法从辣椒疫霉基因组内克隆了漆酶基因Pclac3,该基因全长1 881个核苷酸,编码626个氨基酸,与已知的真菌漆酶基因具有较高的同源性,并且具备漆酶基因的保守区域。利用PCR将该基因克隆于pPIC9K载体,并转化于毕赤酵母GS115,经过1%甲醇诱导表达获得其异源表达产物。将表达产物进行SDS-PAGE检测,获得分子量大约为90 kDa的特异蛋白。利用ABTS法对Pclac3的表达产物粗酶液进行酶活性分析发现,其活性在第11天时最大,达到45 U/mL。这为研究辣椒疫霉漆酶基因家族及探索卵菌漆酶生物活性及其潜在的应用提供了理论依据。     

低温胁迫下马铃薯的数字基因表达谱分析

以马铃薯品种合作88为材料,利用数字基因表达谱(DGE)技术,对–2℃低温胁迫处理后的马铃薯叶片c DNA文库进行差异基因表达谱分析。结果表明,有28 505个基因受低温胁迫诱导差异表达,其中上调表达基因13 703个,下调表达基因14 802个。GO功能显著性富集分析表明,DEGs主要涉及信号生物代谢过程、氧化还原过程、能量代谢、次生代谢过程以及催化活性。KEGG富集分析表明,上调表达基因主要富集于苯丙烷、光合作用天线蛋白、类胡萝卜素的生物合成、苯丙氨酸代谢及淀粉与蔗糖代谢途径,而下调表达基因主要富集于植物激素信号转导途径。利用实时荧光定量PCR(q RT-PCR)验证4个DEGs在低温胁迫条件下的差异表达,其结果与DGE分析结果基本一致,证实了DGE测序结果的可靠性。     

草莓FaSKP1-1基因的克隆与表达分析

为研究草莓中SCF复合体的功能,以栽培草莓品种‘74’为试材,采用同源克隆的方法分离出2个SKP1基因。这2个基因核苷酸序列全长均为519bp,核苷酸序列相似性为98.46%,氨基酸序列相似性为98.84%,并具有特殊的‘GVDED’尾巴结构,分别命名为FaSKP1-1a和FaSKP1-1b(基因登录号分别为KU975057和KU975058)。RT-PCR和dCAPS分析发现FaSKP1-1a和FaSKP1-1b在草莓根、茎、叶、花托、花粉、花柱、果实中均高表达,在花瓣中表达量低。上述结果表明FaSKP1-1可能在草莓SCF复合体的形成过程中发挥重要作用。     

The Relationship Between Vegetation Characteristics and Altitudes in Transitional Permafrost Zone in Xidatan,Qinghai-Tibetan Plateau

Many studies showed that permafrost has profound influence on alpine ecosystem. However, former researches were mainly focused on typical points by temporal scales. There were few studies about the correlation between vegetation characteristics and different altitudes covering a large region in spatial pattern, especially in transitional permafrost(TP). There were continuous permafrost(CP) discontinuous permafrost(DCP) and seasonal frozen ground(SFG) in this study region. The types of permafrost changed from SFG to DCP, and finally become CP as the altitudes of Xidatan increase. In this paper, 112 845 points interpreted by HJ1-B(environment and disaster monitoring and prediction small satellite constellation), vegetation investigation points, thawing layer thickness research sites, ground temperature and water content observation plots were used to examine the spatial pattern of vegetation which were located in different altitudes in Xidatan, a typical TP region, in Qinghai-Tibetan Plateau. Vegetation characteristics, soil moisture content(SMC) and thaw depths were collected in 15 August to 25 August2012. Characteristics of vegetation were mainly represented by fractional vegetation cover(FVC) derived from the normalized difference vegetation index(NDVI), as well as above ground biomass(AGB). In this paper, we analyzed that the distinction of vegetation characteristics in each range through statistics data. These ranges were divided by varied altitudes. For examples, the ranges were divided into 50 m or 100 m. In this study we use a large area plots method to further discuss the relationship between the features of vegetation and the different regions of permafrost based on altitudes shifts in Xidatan. A diagram described the vegetation characteristics variability with rising altitudes in transitional permafrost region was drawn in this paper. Our results illustrated the FVCs first increased in SFG region and then decreased in DCP zone slowly, and in CP region FVCs soared then dropped dramatically. With the altitudes increased, the curve of FVCs indicated a parabolic distribution except a little difference in the first 200 m range.     

危害沙棘的围连环夜蛾生物学特性及防治

围连环夜蛾在我国多有分布，但看见其幼虫的还是很少，因资料中寄主不祥，笔者经室内外观察。发现其危害多种植物的叶片，更喜欢危害沙棘树的叶片，笔者经2a的室内外观察得知。其生物学特性在辽宁西部每年1代，以蛹越夏及冬。对沙棘树危害较重，对栽植的大果沙棘危害尤为严重，在5a生的沙棘树上有100多头幼虫危害，成虫善飞翔。     

密度与环境因子对蒙古黄芪育苗影响研究

为了明确育苗时密度与环境因子对蒙古黄芪生长特征影响,本试验以蒙古黄芪为研究对象,通过单因素田间随机区组试验,测定蒙古黄芪株高、茎粗、根粗和根长4个指标。结果表明,密度对蒙古黄芪影响最大,其次是湿度,最后是光照和电导率。密度与蒙古黄芪株高、茎粗、根粗呈现负相关性,与根长呈正相关性(P<0.05),密度为400株/m ²时,株高、茎粗及根粗最大,密度为1000株/m ²时根长最大。