F18ac菌毛A亚单位基因的克隆与序列分析

根据已经发表的F18ab菌毛A亚单位(FedA/ab)的基因(fedA/ab)[1],设计一对引物,利用PCR技术从表达F18ac菌毛的大肠杆菌2134P株[2]、8199株[3]、8813株[3]中分别扩增到一段序列,并克隆至pGEM-T载体,获得重组质粒T2134PA、T8199A、T8813A.琼脂糖凝胶电泳、序列测定及分析表明,该3个序列大小均为516bp,与fedA/ab(513bp)具有较高的同源性,分别为96.3%、96.5%、95.9%,推导的Fed/ac氨基酸序列与FedA/ab同源性分别为93.0%、93.6%、92.4%.数据表明该实验所克隆的序列均为F18ac菌毛A亚单位(FedA/ac)的基因(fedA/ac).     

新城疫病毒F48E9感染CEF细胞特异性表达基因的筛选鉴定

分别以NDV F48E9和La Sota株感染鸡胚成纤维细胞,于感染后8 h提取NDV感染CEF细胞总RNA,通过mRNA差异显示技术筛选病毒感染诱导表达的特异性基因.主要方法是先以锚定引物经反转录后,然后以9～10 bp随机引物为上游引物,以锚定引物Oligod(T)18为下游引物,进行PCR扩增,PCR产物采用8%的尿素变性PAGE胶电泳进行分离鉴定.对于特征性差异带进行第二次PCR扩增,克隆后测定其核苷酸序列.结果我们发现数个差异条带,选取差异带在500 bp以内的条带,经测序后显示经NDV F48E9感染后,有一条差异条带特异性表达的基因所编码的蛋白与Receptor(TNFRSF)-interacting serine-threonine kinase 2有47 %同源,是一个新基因,其蛋白功能不明确,有可能和细胞的凋亡有关.     

O型口蹄疫病毒VP1 T细胞与B细胞表位基因双拷贝串联表达产物的免疫应答

以一株O型口蹄疫病毒(FMDV)外壳蛋白VP1基因为模板,合成与细胞免疫及体液免疫相关抗原表位肽基因:21-40肽(20AA)和141-160肽(20AA)基因序列,运用基因工程技术构建了含有串联结构21-40(20AA)～141-160(20AA)～21-40(20AA)～141-160(20AA)的2020-2020VP1融合基因表达载体r2020-2020,转化宿主菌BL21(DE3)RIL后诱导表达,表达产物经SDS-PAGE及Western Blot分析显示重组融合蛋白的分子量约为18Ku.动物实验表明,较小剂量的融合蛋白就能诱导豚鼠产生特异性T淋巴细胞增殖反应及抗FMDV中和抗体,证明该融合蛋白可同时激活细胞免疫及体液免疫反应,具有开发成为抗FMDV疫苗的应用价值.     

Effects of gastrin on expression of cyclooxygenase and growth factors in rat gastric mucosa

AIM: To clarify the effects of gastrin on the expression of cyclooxygenase (COX) and several growth factors in rat gastric mucosa. METHODS: Male Sprague Dawley rats were fasted for 24 hours and subcutaneously injected with saline or gastrin 17 at doses of 1 μg/kg, 10 μg/kg and 100 μg/kg, respectively. The expression of COX-1, COX-2, heparin-binding epidermal growth factor-like growth factor (HB-EGF) and hepatocyte growth factor (HGF) in the gastric mucosa were examined using Western blotting and immunohistochemical staining. Effects of a potent gastrin receptor antagonist YM022 on the expression of COX-1, COX-2, HB-EGF and HGF in gastric mucosa were also evaluated. RESULTS: Gastrin dose-dependently increased the expression of COX-2 and HB-EGF in rat gastric mucosa while the expression of COX-1 and HGF did not change significantly after treatment with gastrin. However, pretreatment with YM022 dose-dependently abolished the up-regulation of COX-2 and HB-EGF expression induced by gastrin. CONCLUSIONS: This study demonstrates that gastrin up-regulates COX-2 and HB-EGF expression in rat gastric mucosa, indicating that COX-2 and HB-EGF are involved in pathogenesis of the gastrin-related gastric mucosal hyperplasia and carcinoma of stomach.     

Butylated hydroxyanisole promotes expression of neural specific genes in mouse fetal liver cells through extracellular signal-regulated kinase signaling pathways

AIM: To study the mechanism of butylated hydroxyanisole-induced neural differentiation of fetal liver Sca-1+ cells in vitro. METHODS: Sca-1+ cells from 14.5-day-old mouse fetal liver were isolated with a magnetic cell sorting kit. Cultured cells pretreated with or without extracellular signal-regulated kinase (MEK1) inhibitor, PD98059, were induced by 200 μmol/L butylated hydroxyanisole (BHA) for 24 hours, and then incubated in serum-free medium. Expression of genes in treated or untreated cells were assayed by Western blotting and RT-PCR. RESULTS: There was low level of neuronfilament-L (NF-L) and brain factor-1 (BF-1) in fetal liver Sca-1+ cells, but no neuronfilament-H (NF-H) and tyrosine hydroxylase (TH) was observed. BHA significantly promoted the expression of neuron-specific NF-L, NF-H, BF-1, and TH in fetal liver Sca-1+ cells. NF-L, NF-H, BF-1 and TH increased by 6.32 fold, 2.73 fold, 3.37 fold and 2.68 fold, respectively (all P<0.01 vs untreated cells). However, the expression of BHA-induced genes were inhibited by ERK kinase inhibitor PD98059 (25 μmol/L). CONCLUSION: BHA induces neural differentiation of fetal liver cells through ERK.     

包膜控释尿素对大棚番茄的增产与品质提高效应

对施用包膜控释尿素和普通尿素的大棚番茄进行了田间生产条件下产量与品质对比分析试验。结果表明:与普通尿素相比施用控释尿素有显著的增产效果。D 60控释处理(控释期60 d)产量最高,为68.85 t/hm2,比对照增产19.39 t/hm2,增产率为39.2%;比尿素增产10.10 t/hm2,增产率为17.2%;D 90(控释期90 d)比对照增产15.37 t/hm2,增产率为31.0%;比尿素增产6.08 t/hm2。增产率为10.3%;D 90与D 60两种控释尿素之间产量差异不显著。控释尿素具有增加单株果数、单果重的效果。D 90与D 60氮素当季利用率比普通尿素分别高65.5%和108.8%。硝酸盐含量,以D 90为最低,仅比对照高4.4%,比普通尿素低56%;D 60比对照高15%,比尿素处理低51.6%;尿素处理比对照高137.3%。V c和氨基酸含量,D 90为最高,分别比对照高1.00倍和2.46倍;比普通尿素处理高22%和100%;D 60与尿素两处理V c含量差异不显著,但氨基酸总量前者却比后者高48%。可溶性糖含量以D 60较高,普通尿素为最低。综合品质指标为:D 90>D 60>U。说明控释尿素具有显著地增产效应和提高蔬菜品质的突出效应,特别在提高蔬菜的安全品质方面贡献最大。综合各项,在番茄生产中以选用控释期为60 d的包膜控释尿素为最好。     

苔藓植物生态学研究的数量分析方法

植被数量分析方法在苔藓植被中的应用起步较晚，仅有20多年的研究历史，但发展却十分迅速。这里介绍了目前苔藓植物生态学研究中应用较为普遍的数量分析方法，并探讨了各自的优缺点。     

苏云金素在水中和植株上的衰减与残留

用ＨＰＬＣ检测了苏云金素在鱼塘水和清水中的衰减速率。清水中该物质的半衰期长达４３３小时；在鱼塘水中苏云素的衰减呈双指数消除模型，开始消除半衰期为２．４３－１０．６８小时，终末消除半衰期为１２８．５７－１４２．０１小时。