Messenger RNA splicing in yeast: clues to why the spliceosome is a ribonucleoprotein

The removal of introns from eukaryotic messenger RNA precursors shares mechanistic characteristics with the self-splicing of certain introns, prompting speculation that the catalytic reactions of nuclear pre-messenger RNA splicing are fundamentally RNA-based. The participation of five small nuclear RNAs (snRNAs) in splicing is now well documented. Genetic analysis in yeast has revealed the requirement, in addition, for several dozen proteins. Some of these are tightly bound to snRNAs to form small nuclear ribonucleoproteins (snRNPs); such proteins may promote interactions between snRNAs or between an snRNA and the intron. Other, non-snRNP proteins appear to associate transiently with the spliceosome. Some of these factors, which include RNA-dependent adenosine triphosphatases, may promote the accurate recognition of introns.     

The Xist RNA gene evolved in eutherians by pseudogenization of a protein-coding gene

The Xist noncoding RNA is the key initiator of the process of X chromosome inactivation in eutherian mammals, but its precise function and origin remain unknown. Although Xist is well conserved among eutherians, until now, no homolog has been identified in other mammals. We show here that Xist evolved, at least partly, from a protein-coding gene and that the loss of protein-coding function of the proto-Xist coincides with the four flanking protein genes becoming pseudogenes. This event occurred after the divergence between eutherians and marsupials, which suggests that mechanisms of dosage compensation have evolved independently in both lineages.     

RNA病毒诱导的烟草WRKY转录因子基因的应急表达

植物和病毒在基因水平上的互作是植物感病和抗病的分子基础。以模式植物烟草及其3种RNA病毒（马铃薯Y型病毒,Potato virus Y,PVY;烟草花叶病毒,Tobacco mosaic virus,TMV;黄瓜花叶病,Cucumber mosaic virus,CMV）为试材,旨在研究受病毒共同诱导、并可能在植物抗病反应中具有重要功能的转录因子基因。试验首先获得10个受病毒诱导的烟草同源基因。在比较了3种病毒侵染、低温和盐害协迫以及逆境信号物质(茉莉酸甲酯与水杨酸)处理对上述基因表达的影响后发现, 3种RNA病毒所诱导的植物基因有所不同。但属于WRKY基因家族的06G基因受3种病毒的共同、高效诱导,其表达水平与叶片发育程度呈负相关。研究结果表明, 该基因在植物病毒病发生及植物抗病分子育种中可能有重要意义。     

绵羊MTNR1a基因mRNA表达量的季节性差异

本试验分别于春分、夏至、秋分和冬至4个节气,随机选择体况良好体重相近的蒙古羊8只,外科手术取下丘脑、垂体组织样品,采用RT-PCR方法对其褪黑素受体MTNR1a基因表达进行半定量分析.结果表明:绵羊MTNR1a基因表达存在明显的季节性节律,下丘脑MTNR1a基因表达量在夏至时最低,秋分时最高,春冬季节相近;垂体MTNR1a基因表达量在秋分时最低,夏至时最高,春冬季节也相近.     

荧光定量RT-PCR检测鸡α干扰素mRNA方法的建立

根据鸡IFN-α基因序列设计引物,建立了定量检测鸡IFN-α基因表达水平的荧光定量RT-PCR(RRT-PCR)方法,并对新城疫病毒(NDV)感染的鸡法氏囊、胸腺中IFN-α mRNA表达水平进行了检测.结果表明,该方法对鸡IFN-α,mRNA的扩增效率为103.99%,线性范围为102～10-9,相关系数为0.996,最低能检出73拷贝/反应;熔解曲线分析RRT-PCR产物在(88.6±0.2)℃出现单特异峰;特异性评价结果显示,本方法只扩增鸡IFN-αmRNA,不与常见禽源病原微生物发生交叉反应.对NDV感染鸡的法氏囊、胸腺中IFN-α mRNA表达水平的定量检测结果表明,IFN-α mRNA在感染后36,48 h显著升高.本研究建立的检测鸡IFN-α mRNA表达水平的RRT-PCR方法具有敏感性高、稳定性好和特异性强的特点,为鸡IFN-α mRNA的精确定量提供了新方法.     

Human hair growth deficiency is linked to a genetic defect in the phospholipase gene LIPH

The molecular mechanisms controlling human hair growth and scalp hair loss are poorly understood. By screening about 350,000 individuals in two populations from the Volga-Ural region of Russia, we identified a gene mutation in families who show an inherited form of hair loss and a hair growth defect. Affected individuals were homozygous for a deletion in the LIPH gene on chromosome 3q27, caused by short interspersed nuclear element-retrotransposon-mediated recombination. The LIPH gene is expressed in hair follicles and encodes a phospholipase called lipase H (alternatively known as membrane-associated phosphatidic acid-selective phospholipase A1alpha), an enzyme that regulates the production of bioactive lipids. These results suggest that lipase H participates in hair growth and development.     

DNA-dependent synthesis of RNA is not implicated in growth response of chick comb to androgens

Local application of actinomycin D does not detectably modify growth response of the chick comb to androgens. The mode of action of androgens on chick comb (a connective-tissue organ) appears to differ from their action on epithelial-tissue organs such as seminal vesicles and prostate.     

Expression of bovine leukemia virus genome is blocked by a nonimmunoglobulin protein in plasma from infected cattle

Plasma of cattle infected with bovine leukemia virus contains a soluble factor that blocks the expression of the viral genome in cultured lymphocytes. The blocking factor is not present in plasma of bovine leukemia virus-free cattle or of cattle infected with common bovine viruses. Blocking of bovine leukemia virus expression by the plasma factor is reversible, and seems to be mediated by a nonimmunoglobulin protein molecule.     

Identification of a chromosome 18q gene that is altered in colorectal cancers

Allelic deletions involving chromosome 18q occur in more than 70 percent of colorectal cancers. Such deletions are thought to signal the existence of a tumor suppressor gene in the affected region, but until now a candidate suppressor gene on this chromosomal arm had not been identified. A contiguous stretch of DNA comprising 370 kilobase pairs (kb) has now been cloned from a region of chromosome 18q suspected to reside near this gene. Potential exons in the 370-kb region were defined by human-rodent sequence identities, and the expression of potential exons was assessed by an "exon-connection" strategy based on the polymerase chain reaction. Expressed exons were used as probes for cDNA screening to obtain clones that encoded a portion of a gene termed DCC; this cDNA was encoded by at least eight exons within the 370-kb genomic region. The predicted amino acid sequence of the cDNA specified a protein with sequence similarity to neural cell adhesion molecules and other related cell surface glycoproteins. While the DCC gene was expressed in most normal tissues, including colonic mucosa, its expression was greatly reduced or absent in most colorectal carcinomas tested. Somatic mutations within the DCC gene observed in colorectal cancers included a homozygous deletion of the 5' end of the gene, a point mutation within one of the introns, and ten examples of DNA insertions within a 0.17-kb fragment immediately downstream of one of the exons. The DCC gene may play a role in the pathogenesis of human colorectal neoplasia, perhaps through alteration of the normal cell-cell interactions controlling growth.     

Immunity and intracellular infection: resistance to bacteria in mice infected with a protozoan

Mice infected with the intracellular parasite Toxoplasma gondii for periods of as long as 7 months were resistant to challenge with numbers of Listeria monocytogenes and Salmonella typhimurium that were uniformly lethal to normal mice. This resistance did not appear to depend on the strain of toxoplasma employed or the route of inoculation of either toxoplasma or bacteria. Onset of immunity to listeria was demonstrable as early as 1 to 2 days after infection with toxoplasma. Resistance to toxoplasma was not demonstrable in mice immune to listeria. Interferon did not appear to be a mediator of the immunity observed in toxoplasma-infected mice.     

猪重要性状全基因组关联分析的研究进展

目的揭示大白猪丝氨酸/苏氨酸激酶32B (serine/threonine kinase 32B,STK32B) 基因外显子8多态性及其与母猪繁殖性状的关联。方法采用PCR产物直接测序法检测476头大白猪STK32B基因外显子8区域的SNP位点,结合2177窝母猪繁殖性能记录,采用最小二乘模型分析各SNP位点不同基因型及其单倍型组合对5个繁殖性状的影响。结果大白猪STK32B基因外显子8区域共检出3个SNP位点,包括2个同义突变位点 (A669G、C732T) 和1个错义突变位点 (C749T),分别以AA、CC、CC基因型和A、C、C等位基因的频率最高,3个位点的多态信息含量 (0.1450~0.1744) 和杂合度 (0.1574~0.1930) 总体不高。A669G位点的AA基因型可显著提高仔猪初生窝质量 (P<0.05);C732T位点与总产仔数、产活仔数、仔猪初生窝质量显著关联,其中TT基因型可显著提高总产仔数 (P<0.05),CT基因型可显著提高产活仔数和仔猪初生窝质量 (P<0.05);C749T位点的TT基因型可显著提高仔猪初生窝质量(P<0.05);3个位点单倍型组合ACC/ATC具有极显著提高总产仔数和产活仔数的效应 (P<0.01),ACC/ACT组合具有显著或极显著提高仔猪初生窝质量和断奶窝质量的效应 (P<0.05或P<0.01),ACC/GCT组合具有显著提高断奶仔猪数的效应(P<0.05)。结论研究结果初步揭示大白猪STK32B基因外显子8多态性与母猪繁殖性状显著关联,但3个位点单倍型组合的效应具有一定的性状特异性。     

拟黑多刺蚁hsp90基因的RNA干扰及其对EcR和USP基因mRNA表达的影响

【目的】对拟黑多刺蚁(Polyrhachis vicina Roger)成虫热激蛋白90(Heat shock protein 90,HSP90)基因进行RNA干扰(RNAi),为分析hsp90基因的功能及将RNAi技术用于害虫防治提供参考。【方法】采用Trizol法提取拟黑多刺蚁总RNA,反转录合成cDNA第一链用于hsp90基因的克隆。利用T7RiboMAXTM Express RNAi System将hsp90合成双链RNA(double-stranded RNA,dsRNA),将其配制成120,60,30,15和7.5ng/μL的溶液饲喂拟黑多刺蚁雌蚁、雄蚁和工蚁成虫进行RNAi,共饲喂14d,记录成虫死亡率,确定dsRNA最适质量浓度,并以此剂量对各品级成虫进行干扰,通过荧光实时定量PCR,检测干扰效果及干扰hsp90基因对EcR、USP mRNA表达的影响。【结果】成功获得拟黑多刺蚁hsp90基因片段,其长度523bp,并合成了dsRNA。饲喂dsRNA后发现,dsRNA质量浓度越大,拟黑多刺蚁死亡率越高,在dsRNA质量浓度为120ng/μL时,饲喂5d后成虫全部死亡;30ng/μL是dsRNA干扰的最适质量浓度。以30ng/μL dsRNA干扰后,荧光实时定量PCR结果表明,拟黑多刺蚁每个品级成虫hsp90基因mRNA均被沉默。hsp90基因被干扰后,EcR的表达没有显著变化,USP的表达显著降低。【结论】采用饲喂法成功干扰了拟黑多刺蚁hsp90基因mRNA的表达;hsp90基因与USP基因表达有一定的相关性。