苦瓜MAP30基因的克隆与载体构建

首先提取苦瓜的基因组DNA,然后根据Genbank中已公开发表的MAP30序列,设计一对特异引物MAP301,MAP302,并增添特异定位于内质网的小肽Kozak序列。采用PCR技术,从苦瓜的基因组DNA中扩增出一分子量为800bp左右的片段,回收克隆测序,结果表明,克隆获得片段与已公开发表的MAP30序列同源性达100%,将阳性克隆进行酶切,获得的目标片段与用同样酶切所获得的植物表达载体pEV1和pEV2大片段连接,酶切鉴定重组子构建成特异表达于果实的载体。     

雄蚕品种秋华×平30在山西省的饲养试验

2012年秋季进行了雄蚕品种秋华×平30的饲养试验,结果表明:该品种蚕期发育齐一、眠起齐、抗性强,具有茧层厚、出丝率高、丝质优的特点。     

壬菌铜30%微乳剂防治葡萄霜霉病田间药效试验

试验表明壬菌铜30%微乳剂对葡萄霜霉病有很好的防治效果,使用浓度为200、300mg/kg时,对葡萄霜霉病的防治效果分别为79.9%、85.3%,与氢氧化铜46%水分散粒剂230mg/kg、烯酰吗啉40%悬浮剂250mg/kg处理的防效相当,且耐雨水冲刷,持效期长,价格低廉,值得推广使用。     

浙江选育蚕新品种在陕川应用综述

介绍了陕西安康繁育推广“明丰×春玉”,四川绵阳推广“秋华〈平30”的经验,提出了加快新品种繁育推广的建议。     

一株类NADC30猪繁殖与呼吸综合征病毒的全基因组序列分析

为监测猪繁殖与呼吸综合征病毒(PRRSV)流行毒株的基因变异情况,采用RT-PCR方法对某猪场疑似患有猪繁殖与呼吸综合征(PRRS)的猪组织样品进行鉴定,测序获得全基因,并对该毒株的NSP2基因缺失特征同源性和遗传进化及重组情况进行分析。结果显示:该猪场感染猪体的病原体为PRRSV,该毒株被命名为180404-2fei;180404-2fei毒株的NSP2区存在131(111+1+19)个氨基酸的不连续缺失,与报道的类NADC30毒株缺失特征一致;180404-2fei的NSP2基因与NADC30毒株核苷酸和氨基酸同源性分别为92.2%和90.1%,180404-2fei毒株的ORF5基因与NADC30的核苷酸及氨基酸序列同源性分别为93.5%和92.0%;遗传进化和重组分析结果表明,180404-2fei毒株属于NADC30-Like亚群,可能是由NADC30与HP-PRRSV重组而来。本研究通过对一株类NADC30 PRRSV的全基因组序列进行分析,为研究PRRSV的遗传变异和PRRS的防控提供了参考依据。     

近30年甘肃省农业需水预测与节水高效农业建设途径

以粮食自给为前提,对甘肃省未来30a的农产品需求及高低节水方案下的农业需水进行了预测。通过对水资源供需平衡进行分析后认为,要使农业用水保障供给,农业生产必须走高效用水的道路。提出了发展节水灌溉、建立旱地高效农业、实行水旱互补、统等发展等农业高效用水思路。     

Effect of DNA methylation of miR-30a-5p promoter region on hepatic injury induced by homocysteine

AIM: To explore the role of DNA methylation of microRNA-30a-5p(miR-30a-5p) promoter region in hepatic injury. METHODS: Four-week-old normal mice and cystathionine β-synthase (CBS) single gene knockout mice were used and divided into normal (CBS^+/+, n=12) group and single gene knockout (CBS^+/-, n=12) group, and the mice were fed with high methionine diet for 8 weeks. HL-7702 hepatic cells were routinely cultured in vitro and divided into control group, homocysteine (Hcy) group and Hcy+5-azacytidne (AZC) group. Serum Hcy, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured by automatic biochemical analyzer. The levels of ALT and AST in the cells culture medium were determined by the microplate method. Hepatic injury in the mice were observed with HE staining. Cell viability staining was used to measure the viability of hepatocytes. RT-qPCR was used to detect the expression of miR-30a-5p in the liver tissues and hepatocytes. The correlation between the expression of miR-30a-5p and serum ALT and AST levels was analyzed by Pearson correlation analysis. DNA methylation level of miR-30a-5p promoter region in the liver tissues and hepatocytes was detected by nested landing methylation-specific PCR (nMS-PCR). RESULTS: Compared with the CBS^+/+ mice, the serum levels of Hcy, ALT and AST in the CBS^+/- mice were significantly increased (P < 0.05). HE staining showed the hepatocyte swelling and nuclear fragmentation and dissolution. The expression level of miR-30a-5p in the liver tissues was decreased (P < 0.01). Besides, the expression level of miR-30a-5p in the mice was negatively correlated with serum ALT and AST levels (r²=0.4557, P=0.0003, r²=0.4626, P=0.0003), and the DNA methylation of miR-30a-5p promoter region was increased (P < 0.01). In the HL-7702 cells, compared with control group,the ALT and AST levels were increased in Hcy group (P < 0.05, P < 0.01), and the cell viability was remarkablely decreased. DNA methylation of miR-30a-5p promoter region was increased (P < 0.01), which decreased after treated the cells with AZC (P < 0.05), while the expression level of miR-30a-5p in the cells was increased (P < 0.05). CONCLUSION: Hypermethylation of miR-30a-5p promoter region may play an important role in hepatic injury.     

八氢番茄红素合成酶(PSY2)基因果实特异表达载体的构建

八氢番茄红素合成酶是番茄红素合成的关键酶,通过PCR法获取PSY2基因和E8启动子序列,将目的基因和E8启动子序列构建到植物表达载体pBI101.2中,构建了果实特异表达启动子的八氢番茄红素合成酶基因植物表达载体。并采用PCR、限制性内切酶酶切和序列测定分析法,对重组质粒进行鉴定。结果表明,番茄果实特异性表达PSY2蛋白的重组质粒构建成功;通过农杆菌直接转化技术将其成功转入转化农杆菌LBA4404、EHA105,为下一步PSY2蛋白在番茄果实中特异表达奠定了基础。     

弓形虫P30基因及其功能

弓形虫 P30基因所编码的蛋白为弓形虫主要表面抗原 SAG1 ,其约占速殖子总蛋白量的 5 %,SAG1对虫体侵入宿主细胞及其毒力具重要性 ,并有高度免疫原性和免疫保护性 ,被用于弓形虫疫苗的研制和弓形虫感染的分子诊断