多花黑麦草品种间杂交F2代分子标记遗传分析

运用序列相关扩增多态性(SRAP)和微卫星标记(SSR)技术对6个多花黑麦草亲本品种及其9个杂交F₂代组合80份材料进行遗传分析。筛选出具有多态性的15对SRAP引物、16对SSR引物,SRAP标记平均每对引物扩增出16.7条带,多态性位点百分率为68.88％。SSR标记平均每对引物扩增出14.75条带,多态性位点百分率为58.05％。通过聚类树状图分析得出结论,杂交F₂代内同一组合基本聚为一类,长江2号×剑宝和长江2号×阿伯德组合除外;父本和母本与其杂交F₂代遗传距离与聚类结果基本一致。同时还发现SRAP标记对杂交F₂代及亲代聚类分析可靠性高于SSR标记。     

柔嫩艾美耳球虫棒状体颈部蛋白2基因真核表达载体的构建及在293T细胞中表达

为构建柔嫩艾美耳球虫棒状体颈部蛋白2(Eimeria tenella rhoptry neck protein 2,EtRON2)基因的重组质粒pCAGGS-EtRON2,并转染293T细胞进行表达,以柔嫩艾美耳球虫孢子化卵囊cDNA为模板,经PCR扩增其核心编码区的一部分,将其克隆至pGEM-Teasy载体,构建pGEM-Teasy-EtRON2质粒,双酶切出目的片段后与相应酶切的真核表达载体pCAGGS连接,构建真核表达质粒pCAGGS-EtRON2。该重组质粒经酶切和测序鉴定后转染293T细胞进行表达,分别用免疫印迹和间接免疫荧光鉴定EtRON2基因的表达情况。所构建的真核表达质粒pCAGGS-EtRON2经过双酶切鉴定,可见一条大小约为1172 bp的目的条带,测序结果与GenBank所登录序列完全一致;免疫印迹实验可见大小约为43 kDa的目的蛋白条带,间接免疫荧光实验可以检测到特异性红色荧光。研究结果表明已成功构建了EtRON2的真核表达质粒pCAGGS-EtRON2,并在真核细胞中获得表达,为深入研究EtRON2的生物学功能奠定了基础。     

磷素营养对星星草幼苗抗碱性的影响

通过以不同水平磷素营养培养星星草幼苗，研究磷素营养对星星草幼苗抗碱性的影响，试验结果表明：在２．０％Ｎａ２ＣＯ２胁迫下星星草苗磷素营养的最适浓度为２．８ｍｍｏｌ·Ｌ＾－１ＫＨ２ＰＯ４，此条件下仅对株高，干重和鲜重的增长有促进作用，而且受害程度最小，对Ｎａ２ＣＯ３胁迫的缓解作用最强，在一定程度上可提高星星草幼苗对Ｎａ２ＣＯ３胁迫的抗性。     

应用促排2号提高母猪产仔性能试验

依据“利用GnRH人工干预和控制母畜排卵以提高其繁殖性能”的技术原理，应用国产GnRH产品促排2号，于母猪配种前0－8h一次肌注200μg，试验结果表明可使母猪本胎窝平产仔总数较未使用促排2号的上胎增加1．83头（P＜0．01），窝平活产仔数增加1．78头（P＜0．01），且本胎使用后对下胎产仔性能无不良影响。     

多角体蛋白基因核苷酸序列对HBsAg(PreS2+S)在家蚕中表达的作用

应用PCR突变的方法 ,在乙肝病毒表面抗原 (HBsAg)前S2序列的 5’端融合了BmNPV多角体蛋白基因5’端的 12个碱基 ,获得了融合乙肝表面抗原中蛋白基因 (HBMp) ;通过同源重组将其插入到BmNPV基因组多角体启动子后 ,构建了重组杆状病毒BmPAK HBMp。用重组病毒BmPAK HBMp和BmPAK HBM(带有非融合乙肝表面抗原中蛋白基因 )感染家蚕细胞及蛹 ,对两种病毒的表达产物用ELISA进行了跟踪检测 ,结果表明融合蛋白比非融合蛋白HBsAg活性提高了 6 0 %～ 80 % ,作为HBsAg构成部分的PreS2抗原性提高了 8～ 10倍 ;并且HBsAg和PreS2 Ag表达的时相曲线不同 ,PreS2 Ag的表达量比HBsAg早 1～ 2d达到最大值。ELISA检测和Westernblotting分析表明 ,多角体基因序列的存在更有利于中蛋白全基因 (PreS2 +S)的表达。     

Major outer membrane proteins of Brucella spp.: past,present and future

The major outer membrane proteins (OMPs) of Brucella spp. were initially identified in the early 1980s and characterised as potential immunogenic and protective antigens. They were classified according to their apparent molecular mass as 36–38 kDa OMPs or group 2 porin proteins and 31–34 and 25–27 kDa OMPs which belong to the group 3 proteins. The genes encoding the group 2 porin proteins were identified in the late 1980s and consist of two genes, omp2a and omp2b, which are closely linked in the Brucella genome, and which share a great degree of identity (>85%). In the 1990s, two genes were identified coding for the group 3 proteins and were named omp25 and omp31. The predicted amino acid sequences of omp25 and omp31 share 34% identity. The recent release of the genome sequence of B. melitensis 16 M has revealed the presence of five additional gene products homologous to Omp25 and Omp31. The use of recombinant protein technology and monoclonal antibodies (MAbs) has shown that the major OMPs appear to be of little relevance as antigens in smooth (S) B. abortus or B. melitensis infections i.e. low or no protective activity in the mouse model of infection and low or no immunogenicity during host infection. However, group 3 proteins, in particular Omp31, appear as immunodominant antigen in the course of rough (R) B. ovis infection in rams and as important protective antigen in the B. ovis mouse model of infection. The major OMP genes display diversity and specific markers have been identified for Brucella species, biovars, and strains, including the recent marine mammal Brucella isolates for which new species names have been proposed. Recently, Omp25 has been shown to be involved in virulence of B. melitensis, B. abortus and B. ovis. Mutants lacking Omp25 are indeed attenuated in animal models of infection, and moreover provide levels of protection similar or better than currently used attenuated vaccine strain B. melitensis Rev.1. Therefore, these mutant strains appear interesting vaccine candidates for the future. The other group 3 proteins identified in the genome merit also further investigation related to the development of new vaccines.     

规模化猪场猪圆环病毒2型感染的流行病学调查

20 0 1年 11月至 2 0 0 2年 8月 ,对北京、天津、广东、深圳、山东、山西等地 12个规模化猪场猪圆环病毒 2型 (PCV2 )感染发病群猪进行了临床发病情况调查 ,并采用 PCR方法对所收集的 5 5份组织病料进行 PCV2的检测 ,结果表明 ,12个猪场中有 11个猪场发病猪群表现为断奶后多系统衰竭综合征 ,1个猪场表现为皮炎和肾病综合征。由此可见 ,PCV2感染在我国规模化猪场已普遍存在     

ABA， H2O2与NO对木枣果实成熟生理的影响

在木枣白熟期分别用ABA、H2O2与NO供体硝普纳（SNP）喷洒果实,对其调控木枣果实成熟的生理基础进行了研究。结果显示用不同浓度的ABA,H2O2与SNP处理,枣果实中丙二醛（MDA）的含量明显升高,其中100 mg?L-1浓度时MDA含量为最高;Ca、Vc和β-胡萝卜素含量呈现先降低后升高再下降的趋势,其中75 mg?L-1浓度时Vc和β-胡萝卜素含量最高,50 mg?L-1时Ca含量最高;CAT和APX活性的变化呈现先升高后降低的趋势,75 mg?L-1时CAT和APX活性均最大;DNA含量的变化呈现先缓慢降低后升高再下降的趋势,ABA比H2O2、SNP处理对DNA含量影响更明显,100 mg?L-1时DNA含量最高。结论：ABA、H2O2与SNP对木枣果实发育成熟的影响效果较为明显,其生理效果在多方面表现基本一致,有加快木枣果实成熟衰老的作用。