检测猪流行性腹泻病毒的R-PCR方法的建立

根据猪流行性腹泻病毒 (PEDV)的N基因自行设计和合成了一对可扩增长度为 641bp目的片段的引物 ,成功地建立了检测的猪流行性腹泻病毒的RT PCR方法。对猪轮状病毒 (PRV)、猪传染性胃肠炎病毒 (TGEV)的RT PCR检测结果均呈阴性。对PEDV JS株的RT PCR产物的序列分析表明 ,与CV777株的同源性为 97 3 %。     

君子兰转化酶基因片段(CMCW1)的克隆和序列分析

 分析植物酸性转化酶基因的保守区序列，设计一对PCR引物，以君子兰基因组DNA为模板，采用PCR方法扩增出长约500 bp的DNA片段，克隆入pGEM-TEasy载体，测序结果表明获得君子兰酸性转化酶基因家族的一个成员CMCW1，该基因片段长518 bp，不含内含子，编码172个氨基酸。其序列已在GenBank中登记(登记号为AY151269)。在GenBank中进行同源性检索的结果表明，该成员编码的氨基酸与其它植物细胞壁酸性转化酶编码的氨基酸同源性较高。     

Model establishment and biological behaviour observation of mouse blastocyst co-cultured with hepatocarcinoma cell lines with differently invasive and metastatic potential in vitro

AIM: To explore interaction and biological behaviour changes of two kinds of cells-blastocysts and hepatocarcinoma cells in the same microenvironment. METHODS:The models of mouse blastocysts co-cultured with human hepatocarcinoma cell lines were established, then biological behaviours and mutual effects of the two kinds of cells in co-culture system were observed. RESULTS: Compared with control group, hepatocarcinoma cells with differently invasive and metastatic potential significantly enhanced the rates of blastocyst hatchment , attachment and outgrowth(P＜0.05). There was no significant difference in those among hepatocarcinoma cells co-cultured groups (P＞0.05). The blastocyst hatched and attached to hepatocarcinoma cells with differently invasive and metastatic potential. Then, differential trophoblasts invaded hepatocarcinoma cells. The clear-cut interfaces were gradually formed between both sides. Hepatocarcinoma cells on interface showed changes of growth direction and cell shapes and did not invade blastocysts. CONCLUSIONS: Hepatocarcinoma cells promoted blastocyst development. Blastocysts implanted and invaded hepatocarcinoma cells with differently invasive and metastatic potential in vitro, which indicate that blastocyst implantation in vitro does not relate with the kinds and differential level of interactional cells and the low selectivity maybe relate with high adaptability of early life.     

桃树根癌病防治技术探讨

桃树根癌病是公认最难防治的土传细菌病害之一。在对该病进行较系统研究的基础上,采用多种抗菌药剂、多种方法进行了室内、外药敏测定与防治试验,显示出异菌氰、次氯酸钠、乙蒜素、妥布霉素、环丙沙星、庆大霉素及其混用对防治桃树根癌病菌有明显防效。     

一起未经检疫调运牲畜引发疫情的调查处理

1案情2003年3月23日锡盟正镶白旗改良站从通辽市科左后旗查金台牧场购入26头西门塔尔牛 (其中3头种公牛 ) ,用汽车从科尔沁区运抵锡盟正镶白旗羊场 ,并与2002年11月从通辽科左后旗查金台牧场购入的10头牛混群饲养。24日下午起 ,新购进牛陆续发病 ,至31日病牛已达26头 (新购和原购的牛都有 ) ,旗兽医站接到疫情报告后即派兽医人员到羊场调查情况 ,并采取了相应措施。兽医人员初步诊断为某一类病 ,于是向旗、盟畜牧局和政府上报疫情。并采取病料送自治区兽医站检验诊断 ,确诊为某一类传染病。2处罚经自治区动物防疫监督检验所调查取证 ,这是一…     

表达H3N2亚型猪流感病毒HA基因重组伪狂犬病病毒的构建

将SV40启动子控制下的LacZ基因表达盒和CMV启动子控制下的H3N2亚型猪流感病毒（SIV H3N2）的HA基因插入到伪狂犬病病毒（PRV）通用转移载体pBdTK-Uni中，获得转移载体pLTK-HA。将该载体与PRV Bartha-K61株基因组DNA通过脂质体法共转染Vero细胞，经过10代蓝斑筛选、纯化和PCR鉴定获得了一株插入SIV HA基因的重组伪狂犬病病毒，命名为rPRV-HA。Western blotting和间接免疫荧光试验证实HA基因在重组病毒感染的细胞中获得了表达。用不同的细胞（PK-15、IBRS-2、Vero和鸡胚成纤维细胞）对该重组病毒与亲本病毒的增殖滴度和致细胞病变进行比较，未见显著差异，对第30代重组病毒的HA基因进行序列分析，表明该重组病毒遗传性状稳定。     

Multimarker and rare variants genomewide association studies for bone weight in Simmental cattle

Bone weight, defined as the total weight of the bones in all the forequarter and hindquarter joints, can reflect somebody conformation traits and skeletal diseases. To gain a better understanding of the genetic determinants of bone weight, we used a composite strategy including multimarker and rare‐marker association to perform genomewide association studies (GWAS) for that character in Simmental cattle. Our strategy consisted of three models: (i) A traditional linear mixed model (LMM) was applied (Q+K‐LMM); (ii) single nucleotide polymorphisms (SNPs) with p‐values less than .05 from the LMM were selected to undergo the least absolute shrinkage and selector operator (Lasso) in the second stage (LMM‐Lasso); (iii) genes containing two or more rare SNPs were examined by performing the sequence kernel association test (gene‐based SKAT). A total of 1,225 cattle were genotyped with an Illumina BovineHD BeadChip containing 770,000 SNPs. After the quality‐control procedures, 1,217 individuals with 608,696 common SNPs and 105,787 rare SNPs (with 0.001 < minor allele frequency [MAF] <0.05) remained in the sample for analysis. A traditional LMM successfully mapped three genes associated with bone weight, while LMM‐Lasso identified nine genes, which included all genes found by traditional LMM. Only a single gene, EPHB3, surpassed the significance threshold after Bonferroni correction in gene‐based SKAT. In conclusion, based on functional annotation and results from previous endeavours, we believe that LCORL, RIMS2, LAP3, PRKAR2B, CHSY1, MAP2K6 and EPHB3 are candidate genes for bone weight. In general, such a comprehensive strategy for GWAS may be useful for researchers seeking to probe the full genetic architecture underlying economic traits in livestock.     

补饲水平对甘肃高山细毛羊生产性能的影响

为了探讨补饲水平对高寒牧区甘肃高山细毛羊生产性能的影响,本研究选择12月龄甘肃高山细毛后备母羊48只,随机分为补饲Ⅰ组(颗粒饲料)、Ⅱ组[玉米(Zea mays)和苜蓿(Medicago sativa)干草]、Ⅲ组[玉米、苜蓿干草和燕麦(Avena sativa)干草]和对照组(不补饲),进行冬春季"放牧"+"补饲"试验,比较了后备母羊体重,剪毛量,毛纤维长度、细度、白度、强度和伸度等物理指标。结果表明,试验Ⅰ组、Ⅱ组和Ⅲ组试验羊平均日增重分别比对照组高148.29%、133.09%、96.76%;剪毛量各组之间差异不显著(P0.05);自然长度和伸直长度对照组均极显著低于Ⅰ、Ⅱ、Ⅲ组(P0.01),且自然长度Ⅰ组极显著高于Ⅱ组和Ⅲ组(P0.01),伸直长度Ⅲ组极显著低于Ⅰ组和Ⅱ组(P0.01),Ⅱ组显著高于Ⅰ组(P0.05);细度(直径)Ⅰ组最大,Ⅰ组极显著高于对照组(P0.01),且显著高于Ⅱ组(P0.05);白度Ⅰ组最高,Ⅰ组显著高于对照组(P0.05),其他各组之间均差异不显著(P0.05);强度和伸度补饲组均极显著高于对照组(P0.01),且强度Ⅰ组极显著高于Ⅱ组(P0.01),伸度Ⅰ组显著高于Ⅱ组(P0.05)。由此可见,补饲对体重、羊毛品质都有显著正向影响,高寒牧区冬春季节牧草营养缺乏,为了满足羊只营养需要,使甘肃高山细毛羊后备母羊能在1.5岁进行配种繁殖,在冷季放牧的基础上补饲1号料为最佳选择。     

紫花苜蓿细胞质雄性不育恢复基因的初步定位

以紫花苜蓿(Medicago sativa)不育系MS-GN-1A为母本,恢复系MS178为父本组合构建BSA分离群体,获得的F1代均表现为雄性可育,在大田种植了221株F2代群体单株,盛花期将花粉颗粒染色后,显微镜下观察统计出,不育的F2代株数为57,可育F2代株数164,并没有观察到半不育植株。将所有植株划分为可育和不育两组,并构建可育和不育DNA混池,混池DNA从可育和不育植株组DNA中各随机抽取20个样品,以此对恢复基因定位。随机挑选160对已知的四倍体苜蓿SSR引物扩增基因池DNA,获得2个具有多态性的分子标记,分别是Mt2c12、AW166,初步定位Rf基因在类群Composite5上。将类群Composite5上的所有引物进行合成,进一步进行引物筛选,最终获得4个具有多态性的标记BI68、Mt2c12、BG267和AW776153,遗传距离分别为19.0、20.9、44.6和72.1cM。