RNA干扰技术的应用研究进展

RNA干扰是指由双链RNA(double-stranded RNA,dsRNA)启动的序列特异的转录后基因沉默现象,广泛存在于真菌、植物和动物中。RNA干扰技术是近年来迅速发展起来的高效、特异、易操作的基因阻断技术,在功能基因组的研究中有着广阔的应用前景。文章对RNA干扰的分子机制、生物学功能和RNA扰技术的优势以及其在功能基因组、基因治疗、转基因动物研究、药物开发等方面的应用做一综述。     

猪繁殖与呼吸综合征病毒强毒株HUB2株全基因组序列分析

从湖北省暴发猪"高热病"的猪场分离出1株猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV),并命名为HUB2株.根据GenBank上已发表的PRRSV全基因序列设计引物进行RTPCR扩增,获得PRRSV HUB2株全基因组cDNA序列.测序结果表明PRRSV HUB2株基因组全长15 320 bp(不包括PolyA尾).分析结果显示该毒株与PRRSV美洲型标准株(VR-2332)和欧洲型标准株(LV)全基因核苷酸同源性分别为89.6%和50.3%.说明HUB2属于美洲型毒株.与VR-2332相比,HUB2株非结构蛋白(Nsp2)存在2处不连续的缺失(共缺失30个氨基酸),其缺失位点位于推定氨基酸序列的第481位和532～560位.此次新出现的强毒株全基因组序列特性的揭示为科学防治猪高致病性蓝耳病奠定了理论基础.     

Sequence variability in the mitochondrial DNA control region of five Sebastes species

ABSTRACT: Sequence variability of the control region of mitochondrial DNA in five Sebastes species ( S. thompsoni , S. joyneri , S. inermis , S. schlegeli and S. owstoni ) was investigated. Of 324 nucleotide sites in the control region of S. thompsoni , 56 sites (17.3%) varied, and all 20 individuals had different haplotypes. The other four species had between five and 17 sites (1.5–5.9%), which was fewer than that observed in S. thompsoni . The nucleotide diversity of S. thompsoni was highest (3.45%) among the five species, and that of S. schlegeli was the lowest (0.19%). The results demonstrated that sequence variability exists in the control region of the five Sebastes fishes investigated, and that sequence data in the control region may be suitable for stock structure analysis. Also, the extent of genetic variablitiy in the control region differed among these species. In particular, the mitochondrial DNA control region in S. thompsoni was characterized by high sequence variability, which may indicate a large effective population size.     

Relatedness structure estimated by microsatellites DNA and mitochondrial DNA polymerase chain reaction–restriction fragment length polymorphisms analyses in the wild population of kuruma prawn Penaeus japonicus

ABSTRACT: The genetic structure and variability of four wild populations of kuruma prawn in Japan were examined by estimating relatedness among individuals. The relatedness was estimated by five microsatellites (MS) DNA markers. Examination of relatedness showed that individuals were related significantly in Kumamoto and Kagoshima. In Kagoshima, some individuals showed full-sib level of relatedness. The analysis of mitochondrial (mt)DNA polymerase chain reaction–restriction fragment length polymorphisms (PCR-RFLP) was also performed, showing that the closely related individuals in Kagoshima tended to share a common haplotype. It is, therefore, supposed that there are many kins in Kumamoto and Kagoshima. However, the heterozygosities and allelic and genotypic frequencies in MS-DNA analysis were not significantly different among the localities. Moreover, the haplotype distributions of mtDNA in Kumamoto and Kagoshima were significantly different from other localities. Thus, it is suggested that no spatial differentiations occurred due to the geographic or historical effects between the localities and that there is the possibility of a mixture of hatchery populations in Kumamoto and Kagoshima.     

动物线粒体编码序列相邻碱基组成对核苷酸替换的影响

核苷酸替换偏好性受相邻碱基组成的影响现象已经在核基因组假基因序列和叶绿体基因组的非编码序列和编码序列中发现。通过对动物线粒体基因组编码序列12SrRNA和Cytb基因的核苷酸替换与其相邻碱基组成关系的调查发现,12SrRNA基因序列核苷酸替换中转换和颠换之比（TS／TV）随其相邻碱基A＋T含量的增加而减少,而Cytb基因序列却表现出完全不同的替换类型。     

果树全基因组测序研究进展

 回顾了葡萄、番木瓜、苹果、香蕉、梨、甜橙等10 种果树的全基因组测序的发展历程,基 于全基因组测序结果探讨了不同果树的起源进化,并对果树全基因组测序的后续研究进行了概述,进一 步讨论了果树全基因组测序面临的挑战以及今后的研究重点。     

猪繁殖与呼吸综合征病毒5'非编码区前导转录调控序列及其侧翼序列的结构与功能解析

为解析猪繁殖与呼吸综合征病毒(PRRSV)5'非翻译区(5'UTR)的高级结构与功能,以北美株PRRSV感染性克隆为平台,通过定点突变PCR技术将其5'UTR中一特殊高级调控元件的一级序列进行突变,以改变其二级茎环结构,从而解析该结构的基因调控水平。将突变DNA克隆转染入BHK-21细胞后利用免疫荧光及RT-PCR试验来研究拯救后突变病毒的转录、翻译特性,及通过空斑形态学及病毒生长曲线分析突变病毒的生长特性。结果表明,PRRSV 5'UTR中该高级调控元件的顶端环结构为病毒复制所必需,其只可耐受2个核苷酸的突变;同时发现该调控元件中一保守的茎结构为病毒复制非必需。由此证实了PRRSV 5'UTR中调控病毒复制过程的必需高级结构,为进一步解析PRRSV复制调控元件奠定基础。