生物信息学技术克隆并分析新基因STRF7

为进一步研究信号转导相关的新基因片段BE644250,采用生物信息学方法克隆基全长cDNA,并分析了其ORF,电子表达谱,染色体定位等,之后对全长序列进行了实验验证。电子延伸（contig)获得了729bp的延伸产物,含一个典型的74aa的ORF,命名为STRF7。与已知蛋白无明显同源性,部分地相似于人的源框蛋白CDX－4和酵母的转录调节子ADR6,属一新发现的基因;RT－PCR从IL－6刺激后的U937中克隆了STRF7基因,基序列与电子延伸结果安全一致,进一步的分析显示STRF7在多种组织中表达并定位于第6号染色体上,上述结果显示,STRF7是一个新基因,编码含74aa的蛋白,并且是一个潜在的转录因子。     

High expression of a rice homeobox inE. coli

Homeotic genes share a characteristic DNA segment, the homeobox, which encodes a defined domain of the homeotic protein-homeodomain which seems to mediate the binding to specific DNA sequences, whereby the homeotic protein exerts a gene regulatory function. In this study, the homeodomain encoded by the OSIH-1 (designated fromOryza sativa-Indica homeobox-1) homeobox of rice was overproduced in a expression vector inE. coli, as a form of inclusion body and analyzed by Western blotting and crossed-immunoreaction. Crossed-immunoreaction studies among OSIH-1, Kn-1 and Quox-1 indicate all three are homologous at the protein level. The OSIH-1 homeodomain is then to identify the homeotic target genes. Foundation item: Supported by the National Natural Science Foundation of China (#39570370) Biography: CHENG Xing-guo (1973-), male, Graduate student.     

乐至黑山羊催乳素受体基因cDNA克隆及序列分析

根据GenBank中绵羊催乳素受体（PRLR）基因序列（AF041257.1）设计1对引物,以乐至黑山羊脑垂体总RNA为模板,通过RT-PCR技术对乐至黑山羊PRLR基因cDNA克隆测序和序列分析.结果表明：扩增出的乐至黑山羊PRLR基因cDNA序列长为1743bp,编码581个氨基酸.乐至黑山羊与绵羊、牦牛、欧洲牛、野猪和人的核苷酸同源性分别为：98.34％、94.67％、94.27％、77.48％、74.33％.以核苷酸序列构建分子进化树,乐至黑山羊先与绵羊聚为一类,再与欧洲牛和牦牛聚为一类,而后与野猪聚为一类,最后与人聚为一类.     

A mouse gene related to Distal-less shows a restricted expression in the developing forebrain.

Many genes known to be involved in embryogenesis and morphogenesis of the fruitfly Drosophila melanogaster encode proteins with a highly conserved region of 60 amino acids called the homeodomain. Mammalian counterparts for most of these genes have been identified, including those homologous to the Drosophila homeotic genes or to genes such as evenskipped, engrailed or caudal. We have isolated a murine homeobox gene that encodes a homeodomain similar to that encoded by the Drosophila Distalless (Dll) gene. Dll has a crucial role in Drosophila limb morphogenesis, partially specifying pattern along the proximo-distal axis of the limb. The murine counterpart is expressed in a restricted region of the developing brain, within the diencephalon and the adjacent telencephalic regions.     

Domain of Ultrabithorax expression in Drosophila visceral mesoderm from autoregulation and exclusion

Domains of differential homeotic gene activity are formed at specific positions along the anteroposterior axis of the early Drosophila embryo. Homeotic genes are required continuously throughout development, so that homeotic gene activity has to be maintained independently of the positional information provided in the early embryo. In the ectoderm, the domains of homeotic gene activity partially overlap, but we have found that in the visceral mesoderm at least three of these genes are expressed in adjacent and mutually exclusive domains. It has been proposed that stable, sharply demarcated domains of this type could be established if a homeotic gene product stimulated its own expression locally and inhibited the expression of other homeotic genes, which Meinhardt has termed autocatalysis and mutual exclusion respectively. Furthermore, autocatalysis of this kind can in principle account for the maintenance of homeotic gene activity throughout development. We find that the unique domain of Ultrabithorax (Ubx) expression in the visceral mesoderm is dependent both on autocatalysis and on an exclusion mechanism: Ubx product is required for its own synthesis, whereas the product of the posteriorly adjacent gene abdominal-A represses Ubx expression.     

Drosophila genes Posterior Sex Combs and Suppressor two of zeste encode proteins with homology to the murine bmi-1 oncogene.

The Polycomb group (Pc-G) genes are needed to maintain expression patterns of the homeotic selector genes of the Antennapedia (Antp-C) and bithorax (bx-C) complexes, and hence for the maintenance of segmental determination. We report the predicted protein sequence of the Pc-G gene Posterior Sex Combs (Psc), and of the neighbouring and related gene Suppressor two of zeste (Su(z)2). Both genes encode large proteins that contain a 200 amino-acid domain identical over 37.4% that is also conserved in the murine oncogene bmi-1. At the amino terminus of this domain is a cysteine-rich sequence that has been proposed as a novel type of zinc finger.     

Tobacco floral homeotic gene homologues: partial sequence and expression pattern

Using PCR approach, three cDNA sequences, NTSQUA4, NTSQUA12 and NTSQUA15, were amplified from first_strand cDNAs of wild tobacco flower buds and identified as homologues for floral homeotic genes. All the three clones contained domains that a floral homeotic gene generally had, i.e. I domain, K domain and C_terminal domain except MADS_ box since the PCR primers were designed beyond this region. In addition, the amino acid sequences of them showed 50%-60% identity (70%-80% similarity) with the known floral organ identity class A gene AP1 and SQUA, possibly indicating that they are class A_like genes. NTSQUA4 and NTSQUA412 shared 95% identity in their amino acid sequence, while NTSQUA415 exhibited only 47% identity as compared with NTSQUA4 and NTSQUA12. Within tobacco flower, NTSQUA4 was expressed in sepals, petals and carpels, but not in stamens, while NTSQUA15 was expressed in every whorl of the flower. The possible functions of these genes are discussed.