Cloning and identification of a novel RNF6 transcriptional splice variant Spg2 in human development

RING finger proteins are zinc finger proteins containing the RING motifs. They act mainly as E3 ubiq-uitin ligases, bind the ubiquitin E2 conjugating enzyme and promote degradation of targeted proteins, Many novel genes have been isolated and differentially expressed in human adult and embryo testis by a testis cDNA-array differential display technique. A novel RING finger cDNA is highly expressed in adult testis and at low level in fetal testis. It was named Spg2. It contains a 2055 nucleotide ORF, en-codes a 685-amino-acid RNF6 protein, and has a RING finger in its C terminal. NCBI Blast shows that the gene is located on chromosome 13 and contains five exons. A multiple tissue expression profile also indicates that it is highly expressed in human testis, so we speculate that it may be associated with human spermatogenesis by virtue of the action of its RING domain.     

The structure, expression and function prediction of DAZAP2, a down-regulated gene in multiple myeloma

In our previous studies, DAZAP2 gene expression was down-regulated in untreatedpatients of multiple myeloma (MM). For better studying the structure and function of DAZAP2, a full-length cDNA was isolated from mononuclear cells of a normal human bone marrow, sequenced and deposited to Genbank (AY430097). This sequence has an identical ORF (open reading frame) as the NM_014764 from human testis and the D31767 from human cell line KG-1. Phylogenetic analysis and structure prediction reveal that DAZAP2 homologues are highly conserved throughout evolution and share a polyproline region and several potential SH2/SH3 binding sites. DAZAP2 occurs as a single-copy gene with a four-exon organization. We further noticed that the functional DAZAP2 gene is located on Chromosome 12 and its pseudogene gene is on Chromosome 2 with electronic location of human chromosome in Genbank, though no genetic abnormalities of MM have been reported on Chromosome 12. The ORF of human DAZAP2 encodes a 17-kDa protein, which is highly similar to mouse Prtb. The DAZAP2 protein is mainly localized in cytoplasm with a discrete pattern of punctuated distribution. DAZAP2 may associate with carcinogenesis of MM and participate in yet-to-be identified signaling pathways to regulate proliferation and differentiation of plasma cells.     

Molecular characterization of DnaJ 5 homologs in silkworm Bombyx mori and its expression during egg diapause

A comparison of the cDNA sequences （1 056 bp） of Bombyx mori DnaJ 5 homolog with B. mori genome revealed that unlike in other Hsps, it has an intron of 234 bp. The DnaJ 5 homolog contains 351 amino acids, of which 70 contain the conserved DnaJ domain at the N-terminal end. This homolog orB. mori has all desirable functional domains similar to other insects, and the 13 different DnaJ homologs identified in B. mori genome were distributed on different chromosomes. The expressed sequence tag database analysis of Hsp40 gene expression revealed higher expression in wing disc followed by diapause-induced eggs. Microarray analysis revealed higher expression of DnaJ 5 homolog at 18th h after oviposition in diapause-induced eggs. Further validation of DnaJ 5 expression through qPCR in diapause-induced and nondiapause eggs at different time intervals revealed higher expression in diapause eggs at 18 and 24 h after oviposition, which coincided with the expression of Hsp70 as the Hsp 40 is its co-chaperone. This study thus provides an outline of the genome organization of lisp40 gene, and its role in egg diapause induction in B. mori.     

一条KRAB型锌指蛋白全长新基因的克隆与功能初探

从人胎脑cDNA文库中克隆到一条全长的锌指蛋白新基因的cDNA,命名为ZNF303。序列分析表明,ZNF303的C末端含有7个保守的锌指基序,N末端含有一个KRAB(Krüppel?associated box)结构域。利用肝癌组织表达谱基因芯片杂交证明,该基因在肝癌组织中的表达量有明显降低。认为这种降低可能跟肝癌的形成和转移有密切的关系。利用辐射杂交基因定位技术,得出该基因在人类染色体上的位置是19q13.2。 Abstract: We have cloned a full?length novel zinc finger cDNA of the Krüppel family from human fetal brain cDNA library. Sequence analysis indicates that ZNF303 contains 7 highly conserved zinc finger motifs at the C-terminus and a KRAB(Krüppel-associated box)domain at the N-terminus of the deduced rotein.Hybridization using gene chip of hepatic cancer tissue demonstrates the expressive amount in hepatic cancer tissue is lower than control. We hypothecate that the decreasing of expression amount is related to the formation and metastasis of hepatic cancer.Finally,we show that ZNF303 maps on human chromosome 19q13.2 by radiation hybrid.     

Identification and characteristics of a novel E1 like gene nUBE1L in human testis

A gene, presumably involved in spermatogenesis, was identified and characterized by using cDNA microarray. Hybridization intensity was 2.13 fold higher in adult testis than that in fetal testis.The full length of this gene was 4288bp and it encoded a 578 amino acid protein. Conserved structure and amino acid sequence analysis revealed that the protein contained 1 Thif-domain, 2 UBACT-domains,and a functional active site cysteine lay upstream of UBACT domain, all of them also existed in ubiquitin-activating enzyme E1 and E1 like proteins. So we named this gene as a novel ubiquitin-activating enzyme E1 like gene (nUBE1L). Expression profile showed that nUBE1L was predominantly expressed in testis.Comparison of the expression of nUBE1L in different developmental stages of testis indicated that it was highly expressed in adult testis. In conclusion, nUBE1L is a novel human E1 like gene highly expressed inadult testis, which plays key role in ubiquitin system, and accordingly influences spermatogenesis and male fertility.     

基因诱捕技术及基因诱捕数据库

基因诱捕（gene trap）是基于小鼠胚胎干细胞、报道载体（诱捕载体）建立的一种基因突变方法。诱捕载体在整合位点利用内源基因调控元件模仿内源基因表达,使其表达终止,从而可以阐明内源基因的功能。由于诱捕载体及其报道基因的特点,基因诱捕技术可用于高通量生产,便于小鼠基因功能的大规模研究．为各类疾病动物模型的建立奠定良好基础。     

基因打靶突变小鼠与基因功能研究

ES细胞是建立基因打靶突变小鼠的必要条件 ,也可用于制备转基因动物 .基因敲除、精细突变和条件性基因打靶技术建立的基因打靶突变小鼠在人类遗传病机理研究、基因治疗和基因功能研究方面都有着重要作用 .     

类胡萝卜素合成的相关基因及其基因工程

类胡萝卜素具有多种生物功能,尤其在保护人类健康方面起着重要的作用,如它们是合成维生素A的前体,能够增强人体免疫力和具有防癌抗癌的功效。人体自身不能合成类胡萝卜素,必须通过外界摄入;但类胡萝卜素在许多植物中含量较低,并且很难用化学方法合成。随着类胡萝卜素生物合成途径的阐明及其相关基因的克隆,运用基因工程手段调控类胡萝卜素的生物合成已成为可能。本文综述了微生物和高等植物类胡萝卜素生物合成途径中相关基因的克隆,以及运用这些基因通过异源微生物生产类胡萝卜素和提高作物类胡萝卜素含量的基因工程研究进展。     

壳聚糖作为基因治疗载体的研究

本文从壳聚糖－DNA复合物／微球的形成方法和机理,稳定性,转染细胞效率等方面综述了壳聚糖在基因治疗领域目前的研究现状。     

Identifying Gene Interaction Enrichment for Gene Expression Data

Gene set analysis allows the inclusion of knowledge from established gene sets, such as gene pathways, and potentially improves the power of detecting differentially expressed genes. However, conventional methods of gene set analysis focus on gene marginal effects in a gene set, and ignore gene interactions which may contribute to complex human diseases. In this study, we propose a method of gene interaction enrichment analysis, which incorporates knowledge of predefined gene sets (e.g. gene pathways) to identify enriched gene interaction effects on a phenotype of interest. In our proposed method, we also discuss the reduction of irrelevant genes and the extraction of a core set of gene interactions for an identified gene set, which contribute to the statistical variation of a phenotype of interest. The utility of our method is demonstrated through analyses on two publicly available microarray datasets. The results show that our method can identify gene sets that show strong gene interaction enrichments. The enriched gene interactions identified by our method may provide clues to new gene regulation mechanisms related to the studied phenotypes. In summary, our method offers a powerful tool for researchers to exhaustively examine the large numbers of gene interactions associated with complex human diseases, and can be a useful complement to classical gene set analyses which only considers single genes in a gene set.     

杆状病毒p74基因具有种属特异性

选用苜蓿丫纹夜蛾核多角体病毒杆粒 (AcMNPVbacmid)为材料 ,通过在大肠杆菌中利用RecA基因介导的同源重组 ,将其p74基因剔除 ,并精确地用斜纹夜蛾核多角体病毒 (SpltMNPV)的p74基因进行了替换。所构建的重组AcMNPV杆粒在修饰后的p74基因位点中未留下任何有可能影响该基因表达及功能的选择标记 ,SpltMNPV的p74基因直接位于AcMNPVp74基因的启动子控制下。RT PCR显示替换后的p74基因得到了表达。生物测定结果显示 ,重组病毒AcMNPV杆粒 polhSL74无法通过口服方式感染银纹夜蛾幼虫 ,表明杆状病毒p74基因具有种属特异性。     

益生菌的功能基因导入和基因编辑

益生菌已经在临床和食品领域应用多年,其安全性和有效性已经获得人们的认可。随着分子生物学技术的发展,采用益生菌作为载体进行基因导入或基因编辑,这些遗传改造的益生菌一部分已经作为新的药品或疫苗进入到临床应用阶段。携带功能基因的益生菌定殖于肠道进行表达和缓慢释放,这类益生菌作为活体药物获得益生菌和功能基因的双重功效,可用于治疗某些疑难病症。携带蛋白质抗原基因的益生菌定殖于肠道进行表达,可诱导肠道黏膜免疫、细胞免疫和体液免疫,这是一条更安全的口服疫苗途径。成簇的规则间隔短回文重复序列(clustered regularly interspaced short palindromic repeats, CRISPR)及其相关蛋白(CRISPR-associated protein, Cas)以其高效与便捷性推动了益生菌基因编辑的发展。这篇综述介绍了CRISPR-Cas9操作系统在益生菌方面的应用。对传统遗传操作较难的益生菌采用CRISPR-Cas9技术进行基因编辑,使其基因敲除和基因突变,基因敲入和基因调控等更为简单、高效和易操作。这些CRISPR/Cas9、CRISPRa和CRISPRi技术在...