Enhanced expression of cellular receptors for human interferon α on peripheral lymphocytes from patients with down''s syndrome

The sequence of 370 bases at the 5′-end of bovine thyroglobulin mRNA has been determine. A41 base untranslated segment was found preceeding the ATG initiator codon. It is followed by an open reading frame providing the first data on thyroglobulin primary structure. Analysis of the amino acid sequence demonstrated the presence of an 18 residue hydrophobic segment representing a putative signal peptide. Comparison of the amino terminal sequence of thyroglobulin with that of peptides known to contain thyroid hormones [7,8] demonstrated that the first tyrosine in native thyroglobulin is mainly found as thyroxine in the mature iodinated protein [8]. Our results clearly identify the amino-terminal region of thyroglobulin as an important hormonogenic domain of the protein.     

Construction and characterization of a bacterial clone containing the hemagglutinin gene of the WSN strain (HON1) of influenza virus

A synthetic dodecadeoxynucleotide primer has been used to prepare a double-stranded DNA form of the hemagglutinin (HA) gene of a human influenza virus (WSN strain, HON1). This DNA has been inserted in plasmid pBR322 and cloned in bacterial cells. The insert contains nearly the complete hemagglutinin gene. A restriction map of this insert has been determined and structurally important areas of the HA gene have been sequenced. Amino acid sequences of several regions of the HA protein were deduced from the DNA sequences and compared to the known amino acid sequences of other influenza A viruses. WSN HA shows extensive homology to all influenza A viruses in a few regions, namely the first 17 amino acids of the N-terminus of HA1 (N-terminal polypeptide of HA) and the first 24 amino acids of the N-terminus of HA2 (C-terminal polypeptide of HA). The sequence diverges extensively from other influenza A viruses in most other areas. The sequence of WSN virus HA is similar to that of other HON1 viruses with the exception of the C-terminus of the HA1 peptide. The change in this area may contribute to some of the unique properties of WSN virus among the HON1 viruses. In addition, WSN HA contains a 17-amino-acid precursor before the N-terminus of HA1 and a single amino acid, arginine, connecting HA1 and HA2.     

History of gene therapy

Two decades after the initial gene therapy trials and more than 1700 approved clinical trials worldwide we not only have gained much new information and knowledge regarding gene therapy in general, but also learned to understand the concern that has persisted in society. Despite the setbacks gene therapy has faced, success stories have increasingly emerged. Examples for these are the positive recommendation for a gene therapy product (Glybera) by the EMA for approval in the European Union and the positive trials for the treatment of ADA deficiency, SCID-X1 and adrenoleukodystrophy. Nevertheless, our knowledge continues to grow and during the course of time more safety data has become available that helps us to develop better gene therapy approaches. Also, with the increased understanding of molecular medicine, we have been able to develop more specific and efficient gene transfer vectors which are now producing clinical results.     

小菜蛾烟碱型乙酰胆碱受体α亚基cDNA的克隆、序列分析与不同发育阶段表达分析

烟碱型乙酰胆碱受体(nAChR)介导昆虫中枢神经系统中胆碱能突触兴奋性神经递质的快速传递,也是新烟碱类杀虫剂和多杀菌素的作用靶标。本研究利用RT-PCR和RACE技术,克隆了小菜蛾Plutella xylostella nAChR α亚基的一个新基因(Pxα8)的全长cDNA(GenBank登录号为EU914853)。Pxα8的cDNA序列全长1 744 bp,开放阅读框为1 602 bp,编码534个氨基酸,具有nAChR α亚基的典型特征,与其他昆虫nAChR α8亚基具有77%～96%的相似性,与果蝇nAChR β2亚基具有76%的相似性。Pxα8的开放阅读框存在单核苷酸多态性位点,导致多个位点氨基酸的替换。雌性4龄幼虫的多态性位点多于雄性4龄幼虫,而且雌、雄4龄幼虫的多态性位点均不相同。半定量RT-PCR研究结果表明,Pxα8 mRNA在成虫期表达量高于蛹期和4龄幼虫期。本研究结果为进一步研究小菜蛾nAChR 亚基的多样性和对多杀菌素的靶标抗性机制提供重要基础。     

丙型肝炎病毒RNA打点杂交检测方法同RT－PCR方法的比较

采用ＨＣＶ基因组结构区Ｃ区ｃＤＮＡ探针和非结构区ＮＳ３－４区ｃＤＮＡ探针,建立了用打点杂交（ｄｏｔｂｌｏｔｈｙｂｒｉｄｉｚａｔｉｏｎ）检测血清中ＨＣＶＲＮＡ的方法,同采用ＨＣＶ基因组５’端非编码区的一对寡核苷酸引物通过逆转录－聚合酶链式反应（ＲＴ－ＰＣＲ）检测血清中ＨＣＶＲＮＡ的方法相比较,发现两种方法都能快速早期和特异地检出血清中ＨＣＶＲＮＡ,但ＲＴ－ＰＣＲ法敏感性优于ＲＮＡ打点杂交法。对于无血清学指标的慢性ＮＡＮＢ肝炎病人的诊断,可采用这两种方法。这两种方法的敏感性在很大程度上依赖于引物和探针的敏感性,以及ＲＮＡ提取方法。ＲＴ－ＰＣＲ法适用于诊断病毒血症和复制,打点杂交法适用于研究ＨＣＶＲＮＡ量的变化,对治疗的评价,以及为实验筛选较高滴度的ＨＣＶＲＮＡ阳性样本。     

中华蜜蜂mrjp1 cDNA的克隆及其序列分析

构建中华蜜蜂(Apis cerana cerana)8日龄工蜂头部cDNA文库,利用中蜂基因组的mrjp3部分基因片段作为杂交探针,采用DIG标记筛选cDNA文库,获得mrjps阳性克隆120个;对阳性克隆进行PCR扩增和测序,通过NCBI的BLAST序列比对,获得12个与印度蜂(Apis cerana india)、西方蜜蜂(Apis mellifera L.)mrjp1基因同源的中蜂mtjp1 cDNA片段,并进一步对中华蜜蜂mrjp1的cDNA全序列进行测定和分析。序列比对分析表明,东方蜜蜂(Apis cerana)与西方蜜蜂mrjp1的cDNA序列相似性为93．78％,中华蜜蜂与印度蜂的相似性高达99．36％,这一结果从分子水平证实中华蜜蜂与印度蜂有较近的共同祖先,而东方蜜蜂与西方蜜蜂的亲缘关系较远。     

A novel aspartic acid protease gene from pineapple fruit (Ananas comosus): Cloning,characterization and relation to postharvest chilling stress resistance

A full-length cDNA encoding a putative aspartic acid protease (AcAP1) was isolated for the first time from the flesh of pineapple (Ananas comosus) fruit. The deduced sequence of AcAP1 showed all the common features of a typical plant aspartic protease phytepsin precursor. Analysis of AcAP1 gene expression under postharvest chilling treatment in two pineapple varieties differing in their resistance to blackheart development revealed opposite trends. The resistant variety showed an up-regulation of AcAP1 precursor gene expression whereas the susceptible showed a down-regulation in response to postharvest chilling treatment. The same trend was observed regarding specific AP enzyme activity in both varieties. Taken together our results support the involvement of AcAP1 in postharvest chilling stress resistance in pineapple fruits.