免疫剂量和母源抗体对禽流感重组鸡痘病毒活载体疫苗免疫效力的影响

利用表达 H 5亚型禽流感病毒 (AIV)血凝素基因的重组鸡痘病毒 (r FPV- HA)以不同剂量免疫 1日龄 SPF鸡、有或无母源抗体 (FPV、AIV H5)的商品鸡 ,并于免疫后 2 1d利用同亚型 AIV通过肌肉注射进行致死性攻击 ,通过检测免疫后 HI抗体应答、比较攻毒后发病率和死亡率评价免疫剂量和母源抗体对 r FPV- HA免疫效力的影响。结果发现 ,免疫后 2 1d,15 %～ 2 0 %的 SPF鸡和无母源抗体商品鸡可检出 HI抗体 ,而含母源抗体商品鸡检测不到 HI抗体。利用H5亚型 AIV致死性攻击后 ,10 3～ 10 6 PFU的 r FPV- HA可保护 95 %～ 10 0 %的 SPF鸡和无母源抗体商品鸡抵御强毒攻击 ,使之免于发病和死亡 ;而不同剂量 r FPV- HA接种的含母源抗体商品鸡有 80 %～ 90 %发病和死亡。结果表明 ,在较宽的免疫剂量范围内 ,r FPV- HA对 SPF鸡和无母源抗体商品鸡可提供良好的保护 ,显示出一定的应用前景 ;母源抗体影响 r FPV- HA诱导的免疫应答 ,且提高免疫剂量亦不能克服其干扰作用 ,这提示在实际应用中需优化免疫程序 ,避免母源抗体干扰。     

重组逆转录病毒载体的包装

将3.7kb的LacZ基因片段克隆到含标志基因Neo的逆转录病毒载体，pLXSN与pLNCX的多克隆位点，构建了含有LacZ基因的重组逆转录病毒载体pLLSN和pLNCL.在测定了G418对饥装细胞系PA317的最小致剂量后（最小致死剂量为0.3g/L），通过脂质体Lipofectin与磷酸钙2种介质，分别将2种重组逆转录病毒载体导入包装细胞系PA317进行包装，并以0.3g/L，的G418进行筛选，得到多个G418抗性克隆，经扩大培养，传代，分别得到包装pLLSN与pLNCL的2株阳性细胞，电镜下可见阳性细胞的培养上清中具有逆转录病毒形态特征的成熟病毒粒子，本0研究为运用逆转录病毒载体系统，解决转角因效率低的问题奠定了基础。     

Development of Chinese Near-isogenic Line of Rice and Their Differentiating Ability of Pathogenic Races of Blast Fungus

The rice( Oryza L. ) variety, Lijiangxintuanheigu(LTH), is a universally susceptible variety. A set of near-isogenic lines (NILs) of rice which has single resistance gene for each line was developed by use of LTH as recurrent parent and Kusabue,Tsuyuake, K1, PiNo. 4, K60 and BL1, which are from Kiyosawa‘s differential varieties(KDV), as donor parents. Differentiating ability of these NILs to Philippine isolates was compared with that of their donor parents, KDV and IRRI‘ s NILs. The results of comparative studies were as follows: (1) Differentiating ability of Chinese NILs to Philippine isolates was much stronger than that of their donor parents and that of KDV. (2) Chinese NILs had similar ability to differentiate races to IRRI‘s NILs or higher differentiating ability than them. But IRRI‘s NILs almost had no differentiating ability to isolates from japonica rice-growing regions. They can not be used to differentiate races of isolates from japonica ricegrowing regions. Whereas, Chinese NILs can be used to differentiate races of isolates from both japonica ricegrowing regions and indica rice-growing regions. It was concluded that the set of Chinese NILs can be used as international differentials.     

近交系大鼠DNA指纹图与微卫星DNA比较分析

为了建立一种对近交系大鼠遗传物质进行精确可靠、快速简便的监测方法,利用DNA指纹技术对国内6个品系8个近交系大鼠群体进行了DNA多态性的分析,并与PCR扩增微卫星DNA技术进行了比较.其结果显示(1)不同品系之间DNA指纹图差异较大,同一群体不同个体间DNA指纹图带的相似系数和共有带率除SHR(哈)和WKY(哈)小于0.7外,其他均大于0.9.不同地区同一SHR间和WKY间DNA指纹图也存在差异,相同DNA不同次制作的DNA指纹图谱基本一致.(2)不同品系个体间微卫星DNA具有显著多态性;同一群体不同个体之间除SHR(哈)的SMST位点和WKY(哈)的AGT位点出现一定的差异外,其他均没有差异.DNA指纹图能更精确可靠地反映出动物个体间的遗传背景,而微卫星DNA遗传监测方法比较简便快捷.     

Utilization of Aromatic Rice in Improving Grain Quality of Hybrid Rice

ThesuccessfuldevelopmentandcommercialuseofhybridricehasgreatlyraisedriceyieldlevelinChina .However ,ithasbeenagreatchallengeforbreederstoim provethegrainqualityofhybridricewithoutsacrificeofitsyield ,i.e .,todevelophybridricecombinationswithgoodgrainqualityaswellashighyield .AromaticricelikeBasmatiinIndiaandPakistanandJasmineinThailand ,isfamousforitshighqualityandu niquepleasantflavorofaroma ,soitishighlyfavoredbyriceconsumersaroundtheworld .Itmightbeaneffectiveapproachtoimprovethegrainqual…     

表达HAV结构基因的重组腺病毒鉴定及细胞病变分析

利用RT-PCR方法,从HAV-L8株的RNA中扩增出结构基因(vp3 vp1),克隆到穿梭质粒pXCX2NotI上。采用磷酸钙-DNA共沉淀技术,将腺病毒载体pJM17与pXCX2-CMV-HAV共转染293细胞。在光学显微镜下观察到明显的细胞病变(CPE);经RT-PCR、免疫荧光染色和蛋白印迹鉴定证明HAV的结构基因已插入到腺病毒基因组中;在电子显微镜下观察发现有二十面体的病毒颗粒。以上结果表明获得了重组腺病毒rAdHAV,纯化后的rAdHAV滴度为1×109.0TCID50/mL。     

以CP23重组蛋白为抗原建立检测微小隐孢子虫抗体间接ELISA方法的研究

摘要：以在E.coli高效表达的微小隐孢子虫子孢子表面抗原CP23为抗原，以辣根过氧化物酶(HRP)标记的山羊抗鼠IgG为二抗，建立了检测微小隐孢子虫抗体的间接ELISA方法。经检测筛选出最佳反应条件为1μg／孔纯化的E.coli表达的CP23抗原包被酶标板，用10％免血清进行封闭，以正常E.coli裂解上清液稀释待检血清。实验表明应用CP23重组蛋白作为诊断C.parvum抗原具有特异性高、抗原易纯化和成本低等特点。     

Expression of a recombinant crustacean hyperglycemic hormone of the kuruma prawn Penaeus japonicus in methylotrophic yeast Pichia pastoris

ABSTRACT:   Until now, six crustacean hyperglycemic hormones (CHH) designated Pej-SGP-I, -II, -III, -V, -VI and -VII have been characterized in the kuruma prawn Penaeus japonicus . All CHH consist of 72 amino acid residues and have an amidated carboxyl (C)-terminus. In the present study, we expressed Pej-SGP-III in methylotrophic yeast Pichia pastoris in order to obtain a large quantity of recombinant CHH possessing biological activity. A cDNA encoding Pej-SGP-III that had been previously cloned was processed by polymerase chain reaction (PCR) and the resulting product was ligated into an expression vector. Pichia pastoris was transformed with this vector after which a recombinant Pej-SGP-III was expressed having an additional amino acid residue (glycine) at the C-terminus (rPej-SGP-III-Gly), a form considered to be a putative precursor of this hormone. rPej-SGP-III-Gly secreted into the culture medium was purified by reversed-phase HPLC, and amidated using a peptidylglycine alpha-amidating enzyme. The amidated rPej-SGP-III (rPej-SGP-III) showed hyperglycemic activity in in vivo bioassay almost comparable to that of the natural Pej-SGP-III. rPej-SGP-III thus obtained will be a useful tool not only for its physiological study but also for the determination of its 3-D structure.     

西瓜核型雄性不育两用系POD同工酶研究

用PAGE(聚丙烯酰胺凝胶)电泳技术,研究了西瓜核型雄性不育两用系不同器官的POD(过氧化物酶)同工酶酶谱差异。试验结果表明,不育株与可育株主要差异在雄花蕾,表现为不育株较可育株多2条活性强的酶带;其他器官差异小,仅表现为酶谱带强弱不同。并对POD同工酶与雄性不育性的相关性进行了研究。