Full length antigen priming enhances the CTL epitope-based DNA vaccine efficacy

Although CD8⁺ cytotoxic T lymphocyte (CTL) epitope-based DNA vaccination is valuable experience on vaccine research but many attempts are still continued to achieve acceptable protective response. To study the role of full length antigen in CTL epitope immunization, we evaluated cellular immunity of diverse patterns of complete Herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) and the immunodominant CTL epitope (498–505) DNA injection in C57BL/6 mice. Optimal immune response was observed in the group immunized with the full length of gB in the first injection and CTL epitope in the second and third vaccination as assessed by lymphocyte proliferation assay (MTT), cytokine assay (ELISA) and CTL assay. B cell and spatially CD4⁺ T cell epitopes in full length protein might be important for appropriate priming of CTL immune response. These findings may have important implication for the improvement of CTL epitope based DNA vaccine against HSV and other pathogens.     

朱砂叶螨几丁质合成酶基因1全长cDNA的克隆与表达分析

【目的】克隆朱砂叶螨Tetranychus cinnabarinus几丁质合成过程中的关键酶几丁质合成酶基因,并检测该基因在朱砂叶螨生长发育不同阶段的相对表达量。【方法】本研究采用逆转录聚合酶链反应(RT-PCR)以及c DNA末端快速扩增(RACE)技术首次克隆获得朱砂叶螨几丁质合成酶基因1的全长c DNA序列(命名为Tc CHS1,Gen Bank登录号为KM242062),并使用实时荧光定量PCR技术首次检测了Tc CHS1基因在朱砂叶螨生长发育不同阶段的相对表达量。【结果】朱砂叶螨Tc CHS1基因的c DNA全长为4 881 bp,包括198 bp的5'非翻译区(5'-UTR),4 425 bp的开放阅读框(ORF),258 bp的3'非翻译区(3'-UTR),开放阅读框编码1 474个氨基酸,预测其蛋白质分子质量约为168.35 ku,理论等电点为6.26。其包含EDR和QRRRW这2个几丁质合成酶基因的标签序列。氨基酸序列同源性分析结果表明:Tc CHS1与其他昆虫该基因编码蛋白的氨基酸序列相似度在50%左右,与二斑叶螨Tetranychus urticae的氨基酸相似度最高(98%),与西方盲走螨Metaseiulus occidentalis的相似度为55%。分子系统进化的结果也表明Tc CHS1与其他昆虫的CHS1聚在一起,并且和二斑叶螨具有最近的亲缘关系。荧光定量分析表明Tc CHS1基因在朱砂叶螨生长发育的不同阶段(卵、幼螨、第1若螨、第2若螨、雌成螨和雄成螨)均有表达,在卵和雌成螨中的表达量较高,在第2若螨的表达量最低。【结论】Tc CHS1基因可能在朱砂叶螨生长发育过程中具有重要作用。     

Full length Vpu from HIV-1: combining molecular dynamics simulations with NMR spectroscopy

Based on structures made available by solution NMR, molecular models of the protein Vpu from HIV-1 were built and refined by 6 ns MD simulations in a fully hydrated lipid bilayer. Vpu is an 81 amino acid type I integral membrane protein encoded by the human immunodeficiency virus type-1 (HIV-1) and closely related simian immunodeficiency viruses (SIVs). Its role is to amplify viral release. Upon phosphorylation, the cytoplasmic domain adopts a more compact shape with helices 2 and 3 becoming almost parallel to each other. A loss of helicity for several residues belonging to the helices adjacent to both ends of the loop region containing serines 53 and 57 is observed. A fourth helix, present in one of the NMR-based structures of the cytoplasmic domain and located near the C-terminus, is lost upon phosphorylation.     

苏云金杆菌几丁质酶新基因的筛选和全长基因的扩增

以煮沸冻融法制备PCR扩增模板,利用苏云金芽孢杆菌（Bacillus thuringiensis,Bt）几丁质酶基因特异引物进行15个Bt血清变种的扩增分析,获得9个几丁质酶全长基因扩增产物。经克隆和序列测定,从Bt serovar.entomocidus HD109、Bt serovar canadensis HD224、Bt serovar、alesti HD16和Bt serovar.toumanoffi HD201等4个菌株中分离了几丁质酶新基因。     

Full length ryanodine receptor subtype 3 encodes spontaneous calcium oscillations in native duodenal smooth muscle cells

Two isoforms of the ryanodine receptor subtype 3 (RYR3) have been described in smooth muscle. The RYR3 short isoform (RYR3S) negatively regulates the calcium-induced calcium release mechanism encoded by the RYR2, whereas the role of the full length isoform of RYR3 (RYR3L) was still unclear. Here, we describe RYR-dependent spontaneous Ca(2+) oscillations measured in 10% of native duodenum myocytes. We investigated the role of RYR3 isoforms in these spontaneous Ca(2+) signals. Inhibition of RYR3S expression by antisense oligonucleotides revealed that both RYR2 and RYR3L were able to propagate spontaneous Ca(2+) waves that were distinguishable by frequency analysis. When RYR3L expression was inhibited, the spontaneous Ca(2+) oscillations were never observed, indicating that RYR3S inhibited the function of RYR2. RYR2 expression inhibition led to Ca(2+) oscillations identical to those observed in control cells suggesting that RYR3S did not functionally interact with RYR3L. The presence and frequency of RYR3L-dependent Ca(2+) oscillations were dependent on sarcoplasmic reticulum Ca(2+) content as revealed by long-term changes of the extracellular Ca(2+) concentration. Our study shows that, in native duodenal myocytes, the spontaneous Ca(2+) waves are encoded by the RYR3L alone, which activity is regulated by sarcoplasmic reticulum Ca(2+) loading.     

Full length cDNA structure and deduced amino acid sequence of human 3 beta-hydroxy-5-ene steroid dehydrogenase

Polyclonal antibodies raised against 3 beta-hydroxysteroid dehydrogenase isolated from human placenta were used to screen a lambda gt11 expression cDNA library from the same tissue. The protein deduced from cDNA sequences contains 372 amino acids with a calculated mol wt of 42,216. Since 3 beta-hydroxysteroid dehydrogenase is the enzyme catalyzing the formation of all classes of hormonal steroids, the availability of the cDNA encoding this enzyme opens new possibilities for a detailed investigation of the factors regulating the expression and activity of this crucial enzyme in adrenal, gonadal as well as peripheral tissues.     

Research article

Transgenic Research -     

Review article

Pierre L. van den Berghe, THE ETHNIC PHENOMENON, Elsevier, New York and Oxford, 1981, 301 pp. $25.00.     

Review article

Deciphering the Maya Script. David Humiston Kelley. University of Texas Press, Austin, Texas. 1976. xvii + 334 pp. £19.25.