苜蓿银纹夜蛾核多角体病毒在家蚕蛹体内的复制及重组病毒外源基因的表达

家蚕蛹在复眼着色期,经4℃冷藏24小时后,可被Ac NPV(苜蓿银纹夜蛾核多角体病毒)感染。在蛹体内复制出的多角体大小差异较大,且比在昆虫Sf—21细胞中繁殖的Ac NPV和在蚕蛹体内形成的Bm NPV多角体小。在蛹体内繁殖的Ac NPV的游离病毒可以回返感染Sf—21细胞。由蚕蛹内分离的Ac NPV基因组DNA的限制性内切酶图谱与野生型的Ac NPV相同。以上结果证明Ac NPV可以在蚕蛹体内复制。含HBsAg基因(人乙肝表面抗原)的重组Ac NPV亦可感染家蚕蛹,并能正确表达外源基因。此外,还发现化蛹后第2天的家蚕蛹被注射约5×10~5PFU的Ac NPV,可诱导“人工滞育蛹”现象的发生,在25℃经过30天后蛹仍存活,发育停滞在复眼着色前阶段。     

C肉毒杀鼠素在高寒草甸地区灭鼠技术中的应用

综述C肉毒杀鼠素草原灭鼠应用技术,充分肯定了C肉毒杀鼠素毒力强、适口性好、对非靶动物毒性低、作用缓慢、无二次中毒、易降解和适合规模灭鼠的特点,为鼠害防治提供了新的思路。     

香蕉ACC氧化酶基因(MAO3)的克隆及其表达特性分析

 根据同源扩增得到香蕉(Musa acuminata) ACC氧化酶基因(MAO3) 的核心部分, 再通过3'和5'RACE扩增上、下游序列以及Genome-Walker的方法得到启动子部分, 共获得3 718 bp长度的序列。将所得结果进行聚类分析, 发现香蕉中ACC氧化酶的氨基酸序列非常保守, 各序列间的同一性高达99% ,与单子叶植物和双子叶植物中ACC氧化酶的氨基酸序列的同源性在66.7%～71.8%之间。组织原位杂交试验表明MAO3基因的表达具有组织特异性, 初步认为是在韧皮部筛管组织中特异表达。运用实时荧光定量PCR技术, 实时监测到了MAO3基因和香蕉乙烯受体基因ERS2受机械伤诱导的定量变化。     

罗汉果生产技术研究的思路与发展对策

探讨了罗汉果生产技术研究思路，提出选育、引进优良品种，培育茎尖脱毒壮苗，建立种薯基地，采用容器无土栽培、平地栽培、设施栽培，施用天敌真菌、接种VA菌根真菌防治根结线虫等技术建议。     

鸡传染性腔上囊病病毒VP2基因在昆虫细胞中的表达

为了深入研究鸡传染性腔上囊病病毒（IBDV）VP2基因的结构和功能,利用Bac-to-Bac系统研制出含有VP2基因的重组杆状病毒rBac-VP2,将rBac-VP2感染Sf9细胞,并用抗IBDV VP2特异性单克隆抗体经间接免疫荧光试验检测,证实感染重组病毒Sf9细胞能高效表达IBDV VP2基因产物;Western-blotting分析结果表明,VP2基因表达产物的分子质量约为40 ku.     

中国香瓜与菜瓜地方品种资源白粉病抗性评价

春秋2个季节在温室利用风媒接种方法对搜集的102份香瓜和14份菜瓜种质资源进行了白粉病抗性评价,结果鉴定出7份香瓜和1份菜瓜高抗白粉病,其病情指数为零,1份菜瓜抗病,其病情指数0.9,表明我国香瓜和菜瓜地方品种资源中蕴涵着潜在的改良甜瓜白粉病抗性的基因资源。     

and laminin on adhesion and proliferation of human hepatocellular carcinoma cell

AIM:To explore the effects of PMA(phorbol-12-myristate-13-acetate, a tumor promoter, mimicking the action of diacylglycerol on PKC)and laminin on the adhesion and the proliferation of human hepatocellular carcinoma cells, and provide a new clue to liver cancer treatment.METHODS:Human hepatocellular carcinoma cell line(BEL-7402)was used to identify the endogenous laminin and protein kinase C-α(PKC-α) expression, and the effects of laminin and PMA on the adhesion and the proliferation were also investigatedin vitro.RESULTS:By the effect of exogenous laminin, human hepatocellular carcinoma cell (BEL-7402) possessed endogenous laminin expression and increased the adhesion and the proliferation, which was showed the synergistic action by the effect of PMA in combination. By the action of PMA alone, the proliferation and the PKC-α expression increased by exogenous laminin were decreased, and the adhesion and the endogenous laminin expression were increased.CONCLUSIONS:The finding suggested that the adhesion and the proliferation of human hepatocellular carcinoma cell were closely related to the effects of endogenous or exogenous laminin, which were associated with cPKC-α activity. Therefore, the application of anti-laminin antibody in combination with PKC antagonist might be a new clue to find out the therapy for liver cancer.     

Quick detection of sex determining region protein gene in mouse fetuses

AIM: To detect quickly the Y-chromosome specific sex determining region protein (Sry) gene in mouse fetuses on embryonic day 14.5 with a PCR method. METHODS: We designed specific primers with the OLIGO 5. 0 software. Templates were prepared in 30 minutes by the following way. About 1 mg embryonic tissue but not fetal liver was suspended, and treated with 200μL of lysis buffer, consisting of PCR buffer containing 20 mg/L proteinase K, 0. 5% NP-40, and 0.05% Tween 40, at 60°C for 15 minutes, heated for 5 minutes at 100 °C, 10μL was used as template. The PCR react ion was performed in 50μL, using two sets of primers specific for Sry gene (chromosome Y) and IL-3 gene (chromosome 11) . PCR conditions and cycle numbers were optimized. The assessment of the results was done by electrophoresis in 3% agarose run at high voltage. The specificity of the method was conf irmed by fluorescent in situ hybridization (FISH) using a specific male probe on embryonic tissue cells. RESULTS: Electrophoresis showed that PCR product of male control DNA consisted of a 649 bp product representing the IL-3 gene and a 444 bp product representing the Y-specific Sry gene, female control DNA only one 649 bp product. Fetuses with two bands matching those as seen inmale control DNA are the presumpt ive male fetuses. Fetuses, only the IL-3-associated 649 bp band, are the presumptive female fetuses. These were confirmed by FISH. The ent ire procedure took <3. 5 h. CONCLUSION: The established PCR assay offers a quick, simple, accurate, and sensitive detection of sex determining region protein gene in mouse fetuses. This method allowed the preparation and culture of pure male and female hematopoietic stem cells from fetal tissue.     

Study on relationship between expression of PKCα in glomeruli and development of nephropathy in diabetic rats

AIM:To observe the dynamic changes of expression of PKCα, TGF-β₁ and α-SMA in glomeruli of diabetic rats induced by the alloxon and to invesitigate their roles in the diabetic nephropathy(DN).METHODS:Rats were randomly divided into four groups: normal control group (group A), diabetic group of one week (group B), diabetic group of one month (group C), diabetic group of two months (group D). Immunohistochemistry and Western blotting were used to detect the expression of PKCα, TGF-β₁ and α-SMA in renal tissue of all groups. Blood glucose, triglycerides, cholesterol, creatinine and urine protein were analysed by chemical methods. The morphological changes of renal tissue were checked through microscopy.RESULTS:The expression of PKCα and TGF-β₁ in renal tissue of diabetic groups were increased comparing with those of nomal control group(P<0.05). The mesangial cells expressed α-SMA in two months group. Chronologically the expression of PKCα, TGF-β₁ and α-SMA were positively correlative with each other and the impairment of kidney was also observed.CONCLUSIONS: During the DN process the expression of PKCα increased. PKCα raised GFR and the permeability of glomerular filtration membrane which enhanced urinary albumin excretion. PKCα also increased expression of TGF-β and therefore to induce the expression of α-SMA. The appearance of α-SMA was a marker of the phenotypic transform of renal cells.