Expression of a Modified Cry1Ie Gene in E. coli and in Transgenic Tobacco Confers Resistance to Corn Borer

Gene transfer technology provides an alternativeapproach to breed insect-resistant crops. Insect-resistantgenes from plants or microbes could be introduced intoplants and the expressed insecticidal protein in plantcells could kill the target insects. Transgenic plantsexpressing a corresponding insecticidal crystal protein genefrom Bacillus thuringiensis (Bt) have been developed sincethe early 1980s [1,2]. Analysis of Bt gene sequencesrevealed that they contain numerous motifs seldom foundin p…     

Effect of cDNA fragments in different length derived from Potato Virus Y coat protein gene on the induction of RNA-mediated virus resistance

Potato virus Y (PVY) is a main viral pathogen infecting economic crops such as potato and tobacco plants. Genetic engineering has been so far the most effective method to produce viral resistant plants. Be-cause of the shortage of viral resistant genes in plants, cDNAs derived from viral genes were often used for induction of resistance in transgenic plants (the so- called pathogen-derived resistance)[1]. Among the genes used in the pathogen-derived resistance strategy, the coat protein gen…     

Homology modeling three-dimensional structure of AnxB1 and reducing its immunogenicity by sequence-deleted mutagenesis

We previously isolated and functionally identified a new annexin from Cysticercus cellulosae, which belongs to a novel annexin subfamily and is desig-nated as AnxB1[1]. In the presence of Ca2+, AnxB1 can bind to negatively charged phospholipids like phos-phatidylserine (PS) with high affinity, whereas hardly associates with phosphatidylcholine (PC) even at high Ca2+ concentration. PS is the major component of the platelet membrane and plays a key role in the process of coagulation cascade.…     

In vivo skin penetration and metabolic path of quantum dots

The skin is the largest organ of the body and is a potential route of exposure to sunscreens and cosmetics containing nanoparticles;however,the permeability of the skin to these nanoparticles is currently unknown.In this paper,we studied the transdermal delivery capacity through mouse skin of water-soluble CdSeS quantum dots(QDs) and the deposition of these QDs in the body.QD solution was coated onto the dorsal hairless skin of male ICR mice.Fluorescence microscopy and transmission electron microscopy(TEM) were used to observe the distribution of QDs in the skin and organs,and inductively coupled plasma-mass spectrometry(ICP-MS) was used to measure the 111Cd content to indicate the concentration of QDs in plasma and organs.Experimental results indicate that QDs can penetrate into the dermal layer and are limited to the uppermost stratum corneum layers and the hair follicles.Through blood circulation,QDs deposit mostly in liver and kidney and are difficult to clear.111Cd concentration was greater than 14 ng g-1 in kidney after 120 h after 0.32 nmol QDs was applied to a mouse.These results suggest that QDs have in vivo transdermal delivery capacity through mouse skin and are harmful to the liver and kidney.     

Persistence of VRC01-resistant HIV-1 during antiretroviral therapy

VRC01,a broadly neutralizing monoclonal antibody(bnmAb),can neutralize a diverse array of HIV-1 isolates by mimicking CD4 binding to the envelope glycoprotein gp120.We have previously demonstrated the presence of VRC01-resistant strains in an HIV-1 infected patient during antiretroviral therapy.Here,we report follow-up studies of two subsequent samples from the same patient.With genetic and phenotypic analysis of over 70 full-length molecular clones of the HIV-1 envelope,we show that VRC01-resistant HIV-1 continued to exist and change in its proportion of the infecting virus during treatment with a highly active antiretroviral therapy.Consistent with our previous observation,the resistant phenotype was associated with a single asparagine residue at position 460(N460),a potential N-linked glycosylation site in the V5 region.The persistence and continuing evolution of VRC01-resistant HIV-1 in vivo presents a great challenge to our future preventative and therapeutic interventions based on VRC01.     

Effects of MEK inhibitor U0126 on meiotic progression in mouse oocytes： microtuble organization, asymmetric division and metaphase Ⅱ arrest

In this study we used U0126, a potent and specific inhibitor of MEK, to study the roles of MEK/ERK/p90~(rsk) signaling pathway in the meiotic cell cycle of mouse oocytes. The phosphorylation of MAP kinase and p90~(rsk) in the oocytes treated with 1.5 μM U0126 was the same as that in oocytes cultured in drug-free medium. With 1.5 μM U0126 treatment, the spindles appeared normal as they formed in oocytes, but failed to maintain its structure. Instead, the spindle lost one pole or elongated extraordinarily. After further culture, some oocytes extruded gigantic polar bodies (>30 μm) that later divided into two small ones. Some oocytes underwent symmetric division and produced two equal-size daughter cells in which normal spindles formed. In oocytes with different division patterns, MAP kinase was normally phosphorylated. When the concentration of U0126 was increased to 15 mM, the phosphorylation of both MAPK and p90~(rsk) were inhibited, while symmetric division was decreased. When incubating in medium c     

Expression and Antigenic Characterization of the Epitope-G1 of the Bovine Ephemeral Fever Virus Glycoprotein in Pichia pastoris

The epitope-G1 gene of Bovine ephemeral fever virus(BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1.Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS115.The recombinant P.pastoris strains were selected by a G418 transformation screen and confirmed by PCR.After being induced with methanol,an expressed protein with 26 kDa molecular weight was obtained,which was much bigger than the predicted size(15.54 kDa).Deglycosylation analysis indicated the recombinant G1 was glycosylated.Western blot and ELISA tests,as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity.The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.     

1组曲霉单克隆抗体的制备与鉴定

目的制备并鉴定一组抗曲霉不同抗原的单克隆抗体。方法采用烟曲霉细胞壁抗原成分、分泌抗原和灭活分生孢子,分别免疫BALB／c小鼠,制备单克隆抗体,免疫荧光法鉴定单克隆抗体与曲霉属和念珠菌属抗原的交叉反应。结果获得29株稳定分泌抗曲霉单抗的杂交瘤细胞株,其中用烟曲霉细胞壁抗原成分免疫获得11株,用分泌抗原免疫获得13株,用孢子免疫获得5株;Ig亚类鉴定,11个克隆株为IgG1亚类,3个克隆株为IgG3,15个克隆株为IgM。免疫荧光法鉴定29株单抗特异性识别烟曲霉细胞壁抗原,与其他曲霉抗原有交叉反应。结论29株单克隆抗体,对于建立侵袭性曲霉感染早期诊断方法、筛选曲霉保护性抗体以及研究抗体保护机制奠定了实验基础。     

比喻在细胞生物学教学中的应用

细胞生物学的内容比较抽象,学生听课时往往感到枯燥乏味,作者在教学中对某些内容试用比喻的方式予以表达,这对于活跃课堂气氛、把抽象的内容形象化、增强学生的记忆有一定作用。举例如下:液态镶嵌模型:如果把类脂双分子层比作浩瀚的海洋的话,那么镶嵌蛋白就是海洋中的岛;周围蛋白是飘浮在海洋中的船只。染色质和染色体:染色质和染色体是同一种物质在细胞周期不同时相的不同的表现形态。这就好像水一样,水在零度以下,表现为固态,在零度以上表现力液态或汽态,