五种真蝽(Pentatoma Olivier) mtDNA-COII基因序列分析及系统发育关系初探(半翅目:蝽科)

测定并比较分析了真蝽属(Pentatoma)5种昆虫：褐真蝽P．semiannulata、斜纹真蝽P．illuminata、红足真蝽P．rufipes、H本真蝽P．japonica和金绿真蝽P．metallifera以及益蝽亚科2种外群种类(并蝽Pinthaeus humeralis和蠋蝽Arma custos)mtDNA—COII基因部分序列,依据分子数据建立了该5种昆虫的系统发育关系。结果显示,5种真蝽COII基因A T平均含量(73％)高丁弹尾目、缨尾目和直翅目昆虫,低于双翅目、鳞翅目及膜翅目昆虫,而与缨翅目的蓟马和半翅目的突角长蝽Oncopeltus fasciatus非常接近。系统发育结果显示,红足真蝽与日本真蝽亲缘关系最近,形成一个姊妹群,斜纹真蝽与金绿真蝽亲缘关系较近,也形成一个姊妹群,褐真蝽为较早分化的种类。     

Feeding scallop shell powder induces the expression of uncoupling protein 1 (UCP1) in white adipose tissue of rats

Previously we found that the organic components in scallop shell promote lipolysis in differentiated 3T3-L1 and C3H10T1/2 adipocyte cells, and that incorporating scallop shell powder into the diet of rats reduced the amount of white adipose tissue. In this study, we used RT-PCR to investigate the effect of ingesting scallop shell powder on the gene expression profile of uncoupling proteins (UCPs) regulating energy metabolism in rats.Feeding of scallop shell powder increased mRNA levels of UCP1 and UCP2 in white adipose tissue. By contrast, scallop shell powder had no effect on the expression of UCP1 in brown adipose tissue, although the expression level of UCP2 mRNA decreased significantly. These results suggest that feeding scallop shell powder increases gene expression of UCP1 that may regulate energy metabolism in white adipose tissue, resulting in the observed reduction in weight of white adipose tissue.     

Angioarrestin(hARPl)C端FD区的克隆、表达及其免疫活性鉴定

 Angioarrestin是一种具有潜在应用价值的肿瘤血管形成抑制因子.利用DNA重组法构建了angioarrestin C端 hFD cDNA 和麦芽糖结合蛋白(MBP)重组原核表达质粒 pMAL-C2-hFD.将重组质粒转入大肠杆菌E.coli BL21（DE3）,经0.3 mmol/LIPTG 在37℃条件下诱导表达4h,SDS-PAGE 检测,融合蛋白表达量约占细菌总蛋白的20%.Western印迹证实,目的蛋白N端带有MBP标签.取表达上清纯化、透析、浓缩并冻干,以此为抗原免疫Balb/c小鼠制备多克隆抗体.此多抗可以与pET 22b（+）表达系统获得的 hFD重组蛋白发生良好的抗原抗体反应,ELISA检测多抗效价达1∶10240.实验证明：通过基因重组可获得angioarrestin C端hFD在大肠杆菌中的高效表达蛋白,且该蛋白具有较高的免疫活性.以此为抗原制备的抗angioarrestin多克隆抗体为深入研究angioarrestin提供了材料.     

利用PCR方法鉴定hiTAIL-PCR扩增产物中的质粒骨架片段

T-DNA突变体是研究基因功能的重要资源。高效热不对称交错PCR (hiTAIL-PCR)是克隆突变体中T-DNA插入位点侧翼序列的常用方法。然而我们发现, 利用hiTAIL-PCR克隆到的一些侧翼序列并不对应于宿主的染色体DNA序列, 而是质粒的骨架DNA片段。通过设置1组RB-S4/AC1或者LB-A4/AC1对照反应, 用PCR方法鉴定了hiTAIL-PCR扩增产物中位于T-DNA侧翼的质粒骨架片段。在后续分析中, 通过排除这些片段, 提高了利用hiTAIL-PCR获得宿主染色体DNA片段的效率。同时, 通过调整反应程序, 使得整个PCR的反应时间也大为缩短。在拟南芥(Arabidopsis thaliana) T-DNA突变体drf1侧翼序列的克隆实例中, 对照反应的引入将hiTAIL-PCR中需鉴定的22条扩增产物降至4条, 效率提高了81.8%。     

原生动物伪红色双轴虫细胞纤毛器微管胞器的直接荧光标记

应用荧光紫杉醇直接荧光标记法显示,原生动物纤毛虫伪红色双轴虫(Diaxonellapseudorubra)细胞纤毛器微管中,口围带基部含小膜托架及与托架相联系的肋壁微管;额腹横棘毛基部含前纵微管束、后纵微管束、横微管束和周围微管束,其微管在不同棘毛基部的定向和发达程度不一;缘棘毛基部含前纵微管束、后纵微管束。细胞形态发生过程中,前仔虫口纤毛器微管独立发生于老口围带内侧,在细胞形态发生末期新纤毛器微管形成时,尚有部分老额棘毛、横棘毛和缘棘毛残存,此后老结构逐渐被吸收。结果表明,伪红色双轴虫的纤毛器基部微管的分化很可能具有种属级的特异性,新纤毛器微管分化过程中老结构可能具有定位和物质贡献作用。     

茶碱与胃蛋白酶相互作用的光谱性质研究

采用紫外光谱法和荧光光谱法研究了茶碱与胃蛋白酶的结合作用。观测到茶碱使胃蛋白酶的紫外吸收峰增强,特征荧光峰淬灭。Stern-Volmer淬灭曲线显示,茶碱对胃蛋白酶的荧光淬灭很可能是一个单一的静态淬灭过程。     

Alteration in gene expression profile and biological behavior in human lung cancer cell line NL9980 by nm23-H1 gene silencing

Lung cancer is the leading cause of cancer death in both men and women. Tumor metastasis is an essential aspect of lung cancer progression. nm23-H1 is a metastasis suppressor gene. The molecular mechanism by which nm23-H1 suppresses the metastasis is still unclear. Here, we compared the gene expression profile of human large cell lung cancer cell line NL9980 by nm23-H1 gene silencing with that of negative control cells to comprehensively investigate nm23-H1-mediated changes in gene expression of NL9980 cells. Microarray assay revealed that expression of 733-known genes (1.9%, 733/38,500) were altered in response to nm23-H1 gene silencing, including 466 upregulated genes and 267 downregulated. real-time PCR assay of the expression changes indicated that 81.82% (45/55) of verified genes were consistent with that observed in microarray assay. The upregulated genes included MMP-1, -2, SNAI2, CXCL1, 2, 3, PAI-2, while the downregulated genes included cystatin B, TIMP-2, E-cadherin, centrin-2, all of which have been associated with tumor metastasis. Furthermore, we confirmed by Western blot that the expression of MMP-1 and -2 were significantly increased while that of cystatin B was dramatically decreased in NL9980-nm23-H1 silencing cells. The NL9980-nm23-H1 silencing cells exhibited significantly more S phase growth and invasive ability. Thus, silencing of nm23-H1 gene caused metastasis-related gene expression changes in lung cancer cells. The knockdown of nm23-H1 expression may change the lung cancer cells to a more invasive phenotype through alteration in the expression of a set of genes.     

金色蝗属的分类研究及一新种记述（直翅目: 剑角蝗科）

本文记述采自内蒙古贺兰山地区金色蝗属Chrysacris 1新种, 即白纹金色蝗Chrysacris albonemus Zheng, Zhang et Zeng sp. nov. 该新种近似于山间金色蝗Chrysacris montanis Zhang et Zheng, 1993, 主要区别为: 1)头顶及头部背面具中隆线; 2)前胸背板沟前区长为沟后区长的1.6~1.8倍; 3)前翅前缘脉域宽为中脉域宽的1.5~2倍。附有金色蝗属分种检索表。模式标本保存于陕西师范大学动物研究所昆虫标本室。     

铁线蕨中间纤维的研究及某些植物类角蛋白的比较分析

应用整装电镜制样技术,结合选择性抽提方法在铁线蕨（ＡｄｉａｎｔｕｍｐｈｉｌｉｐｐｅｎｓｅＬ．）叶细胞中观察到直径１０ｎｍ的纤维网络结构。免疫印迹结果显示：经选择性抽提得到的纤维蛋白与动物角蛋白抗体有免疫交叉反应,间接免疫荧光标记也得到类似结果,而且此类蛋白能在体外自组装为１０ｎｍ或更粗的纤维。说明蕨类植物细胞中存在类角蛋白中间纤维网络。免疫印迹结果表明,螺旋藻（ＳｐｉｒｕｌｉｎａｓｕｂｔｉｌｌｉｓｉｎａＫｕｔｚ．）细胞,地钱（ＭａｒｃｈａｎｔｉａｐｏｌｙｍｏｒｐｈａＬ．）叶状体,铁线蕨（Ａ．ｐｈｉｌｉｐｐｅｎｓｅＬ．）、银杏（ＧｉｎｋｇｏｂｉｌｏｂａＬ．）、白菜（ＢｒａｓｉｃａｐｅｋｉｎｅｎｓｉｓＲｕｐｒ．）的叶组织经选择性抽提后得到的蛋白均与动物角蛋白抗体有免疫交叉反应。其中,螺旋藻仅含有两种类酸性角蛋白,而其余４种植物材料均含有３种类酸性角蛋白及３种类碱性角蛋白。结合以往实验结果,我们认为类角蛋白在植物细胞中是普遍存在的。     

Cloning, sequencing and prokaryotic expression of cDNAs for antifreeze protein family from beetle Tenebrio molitor

Partial cDNA sequences coding for antifreeze proteins in Tenebrio molitor were obtained by RT-PCR.Sequence analysis revealed nine putative cDNAs with a high degree of homology to Tenebrio molitor antifreeze protein genes published in GenBank.The recombinant pGEX-4T-l-tmafp-XJ430 was introduced into E.coli BL21 to induce a GST fusion protein by IPTG.SDS-PAGE analysis for the fusion protein shows a band of 38 kDa.pCDNA3- tmafp-XJ430 was injected into mice to generate antiserum which was later detected by indirect ELISA.The titer of the antibody was 1:2000.Western blot-ting analysis shows that the antiserum was specifically against the antifreeze protein.Our results laid the founda-tion for further studies on the properties and functions of insect antifreeze proteins.