荷斯坦乳牛乳腺和乳腺上皮细胞中Toll样受体及辅助因子编码基因的检测

参照牛TLR4、TLR2、CD14、MD-2基因序列设计了相应基因的引物。采用RT-PCR技术检测了体外培养的荷斯坦乳牛乳腺和乳腺上皮细胞中Toll样受体TLR4、TLR2及辅助因子CD14、MD-2基因。结果显示,乳腺上皮细胞中存在TLR4、TLR2、CD14和MD-2四个基因的表达,而乳腺中除MD-2未检测到外,其余3个基因均扩增成功。说明该受体及辅助因子可能参与了乳腺的先天性免疫防御。该研究为探讨乳腺的先天性免疫及乳腺上皮细胞在乳腺先天性免疫中的作用奠定了基础。     

Epidermal growth factor receptor-protein kinase interactions in hepatic membranes of rainbow trout (Oncorhynchus mykiss)

The association between the presence of EGF binding and its associated tyrosine kinase activity was investigated in the hepatic membranes of rainbow trout (Oncorhynchus mykiss). Time course experiments indicted that the binding of EGF to hepatic membranes was temperature dependent and resulted in increased phosphorylation of membrane components. The phosphorylation reaction was dependent on the presence of divalent ions (Mg²⁺ and Mn²⁺) but not monovalent ions (Na⁺ and K⁺) or calcium. Western blot analysis of phosphorylated trout membranes indicated protein bands at 150 and 180 kDa which is similar to that observed in mammals. Use of specific substrates and protein kinase inhibitors also indicated that the increase in EGF stimulated phosphorylation was related to an increase in tyrosine kinase activity. These data support the hypothesis that trout have an EGF-like receptor that has binding and biological activities similar to that observed in mammalian species.     

甘蓝自交不亲和决定因子的体外表达和相互作用的检测

 S位点受体激酶(SRK)和S位点富含半胱氨酸蛋白/S位点蛋白11(SCR/SP11)分别为甘蓝自交不亲和(self-incompatibility, SI)信号传导的雌雄决定因子。为了深入研究两者的作用机理和进行人工调控，本研究以结球甘蓝ZQ为材料，利用pET·NusA融合蛋白表达系统，将包含SRK胞外域和跨膜域的mSRK蛋白和SCR蛋白在大肠杆菌BL21中融合表达，经SDS-PAGE电泳检测表达出融合蛋白大小分别约为116 kD和74 kD。进一步将两融合蛋白进行体外相互作用的检测，结果表明mSRK与SCR蛋白能够相互结合形成稳定的复合体，这为下一步实现SRK-SCR复合体聚合与解离的人为调控提供了技术平台。     

催乳素及其受体对乳腺发育研究进展

本文主要对催乳素受体的结构、作用机理、功能及其在不同时期的乳腺发育的不同作用进行的论述。总结了近几年来的有关研究进展,为进一步的研究工作提供参考资料。     

Adenovirus-mediated mPPARγ1 gene overexpression inhibits IFN-γ-induced galectin-9 gene and protein expression in ECV304

AIM:To investigate whether adenovirus-mediated mPPARγ1 gene overexpression inhibits IFN-γ-induced galectin-9 gene and protein expression in ECV304. METHODS:A replication-deficient recombinant adenovirus expression vector of mPPARγ1 was constructed by using the AdEasy system. ECV304 were incubated for 24 h with 1×104 U/L, 5×104 U/L, 1×105 U/L and 2×105 U/L IFN-γ, respectively. ECV304 stimulated with 1×105 U/L IFN-γ were divided into 4 groups in random: P group (PPARγ1 gene overexpression), T group (treated with troglitazone 40 μmol/L in DMSO), PT group (PPARγ1 gene overexpression+troglitazone treatment) and control group. Changes of PPARγ and galectin-9 in mRNA and protein levels in different groups and subgroups were investigated by RT-PCR and immunoblotting. RESULTS: Galectin-9 expression was very few in normal ECV304. IFN-γ induced the expression of galectin-9 in ECV304. Degree of galectin-9 expression increased with the dose of IFN-γ. PPARγ1 gene overexpression inhibited IFN-γ-induced galectin-9 expression in ECV304. Galectin-9 mRNA and protein expressions from PT group and P group were inhibited in similar degree (P>0.05). However, this effect was not observed in troglitazone intervention (P>0.05). PPARγ expression was also very few in normal ECV304. PPARγ1 gene overexpression/activation had no effect on endogenous mPPARγ expression. CONCLUSION: This may partly contributed to the anti-inflammatory and immuno-regulatory effect of PPARγ1 gene overexpression by inhibiting IFN-γ-induced galectin-9 gene and protein expression in ECV304.     

Leptin长型受体与神经肽Y在猪前脑内共存

用原位杂交与免疫组织化学相结合的方法研究了 5头三元杂交仔猪前脑内leptin长型受体mRNA与神经肽Y(NPY)免疫反应阳性物质的共存关系。实验结果表明 ,leptin长型受体mRNA和NPY免疫反应阳性神经元分布于下丘脑漏斗核和前室周核、海马、杏仁核及大脑皮质。上述结构内一些表达leptin长型受体mRNA的神经元也含有NPY免疫反应阳性物质 ,即leptin长型受体与NPY共存于同一神经元内。提示leptin可能通过下丘脑漏斗核和室周核以及边缘系统的海马和杏仁核 ,甚至大脑皮质中表达leptin长型受体mRNA的NPY免疫反应阳性神经元调节动物的摄食、能量代谢、生长和繁殖等活动。     

小麦转化受体体系的建立--小麦幼胚组织培养研究

建立了一个以小麦幼胚为转化受体,转化后快速成苗的体系.筛选出适合于不同小麦品种的愈伤组织诱导培养基MS+2mg·L-1 2,4-D+0.4mg·L-1 IAA+0.2mg·-1 KT,愈伤诱导率达94.87%～99.04%.幼胚接种后,暗处理与弱光、强光合理配置利于绿芽的快速形成,在分化培养基中添加2.0g·L-1活性炭或1mg·L-1多效唑利于小麦再生苗的正常生长,从而提高移栽成活率.     

84K杨基因转化受体系统的建立

以84K杨为研究对象,通过建立叶片再生系统及卡那霉素敏感性试验,确立了稳定高效的基因转化受体系统。结果表明,以MS培养基为基本培养基,0.5～1.0mg/L6-BA+0.01～0.1mg/LNAA组合时,叶片出芽率不到50%,每叶平均生芽数5～6个;1.0～1.5mg/L6-BA+0.02mg/L2,4-D组合时,叶片出芽率达90%～100%,每叶平均生芽数约20个。用后者附加不同质量浓度梯度卡那霉素,确定叶片外植体芽诱导卡那霉素临界筛选质量浓度为20mg/L。利用对84K杨芽生根效果较好的培养基GMS+0.01mg/LNAA+0.25mg/LIBA,附加不同质量浓度梯度卡那霉素,筛选出芽生根卡那霉素临界质量浓度亦为20mg/L。     

二花脸猪和大白猪睾丸生长激素受体基因表达的发育性变化

随机选取0、3、20、30、90、120和180日龄二花脸公猪和大白公猪各4头，用相对定量RT-PER法研究睾丸组织中生长激素受体（GHR）基因表达的发育性变化。结果表明，二花脸猪与大白猪睾丸GHR mRNA表达的发育模式不同，二花脸猪GHR mRNA相对表达量从0到30日龄逐渐上升，在90和120日龄呈阶段性降低（P〈0．05），之后在180日龄又再次上调；大白猪GHR mRNA相对表达量在出生当日最高，3日龄显著下降（P〈0．05），随后直至180日龄维持在较低水平并相对恒定。总的来说，二花脸猪睾丸GHR mRNA相对表达量显著高于大白猪（P〈0．05）。提示：不同品种猪睾丸GHR mRNA表达具有特定的发育模式，并可能与性成熟的早晚有关。     

褪黑激素受体1A基因外显子2与小尾寒羊高繁殖力的关联

以褪黑激素受体1A(melatonin receptor 1A,MTNR1A)基因为候选基因,通过2种限制性内切酶(MnlⅠ、RsaⅠ),采用PCR-RFLP技术检测MTNR1A基因第2外显子主要序列在高繁殖力绵羊品种小尾寒羊中的单核苷酸多态性,同时研究该基因对小尾寒羊高繁殖力的影响.结果表明,MTNR1A基因外显子2的605位碱基处表现出MnlⅠ酶切多态性,在小尾寒羊中检测到3种基因型:MM(236 bp/236 bp)、Mm(236 bp/303 bp)、mm(303 bp/303 bp),MM和Mm基因型频率明显高于mm基因型频率.MTNR1A基因外显子2的604位碱基处表现出RsaⅠ酶切多态性,在小尾寒羊中检测到3种基因型:RR(267 bp/267 bp)、Rr(267 bp/290 bp)、rr(290 bp/290 bp),RR和Rr基因型频率明显高于rr基因型频率.复合基因型MMRR、MMRr、MmRR、MmRr的频率明显高于MMrr、mmRR、mmRr的频率,没有检测到mmrr复合基因型.小尾寒羊产羔数在MnlⅠ和RsaⅠ酶切复合基因型之间没有显著差异,但至少携带一个拷贝等位基因M的小尾寒羊比不携带M的具有稍微较高的产羔数,基因型为mm的小尾寒羊产羔数比其他基因型的小尾寒羊平均低0.26只.