江苏汉族人生长激素受体基因多态性研究

为了解生长激素受体(GHR)基因多态性在江苏汉族人群中的分布,通过多重PCR及PCR-RFLP的方法对269例江苏地区正常汉族人进行GHR基因3个位点的多态性分析,并与文献中报道的其它种族和地区的人群进行比较分析.结果显示江苏地区汉族正常人群中GHR基因型以fl/fl、I/L、P/P发生频率最高,这种分布在男女之间没有差异,与其它地区汉族人比较没有明显差异,与日本人差异也不明显,而与法国、俄罗斯人差异明显.研究表明,GHR基因3个位点多态性在不同种族之间分布存在着明显的差异,这种差异有可能是导致不同种族间人体测量学差异及对一些疾病的易感性及对药物的反应性不同的因素之一.     

重组人脂联素基因构建、表达、纯化及活性鉴定

根据脂联素cDNA序列设计并合成9条寡聚核苷酸片段,经重叠PCR技术获得重组人脂联素基因.构建pET-22b( )/ADPN表达载体,转化大肠杆菌BL21(DE3),IPTG诱导表达.目的蛋白以包涵体的形式表达,表达量占菌体总蛋白的30%以上.表达产物通过Ni-NTA柱纯化,SDS-PAGE分析显示,其分子量约为30 ku,与预期结果一致.内皮细胞生长抑制实验证实,重组人脂联素生理浓度下具有抑制人内皮细胞生长的活性并呈时间依赖性.以上工作为进一步研究脂联素的生物学功能奠定了基础.     

水稻16kDa醇溶蛋白启动子克隆及载体构建

水稻种子16 kDa醇溶蛋白是水稻种子成熟过程中,由16 kDa基因编码,16 kDa醇溶蛋白启动子调控,在胚乳中特异表达的蛋白质.以水稻基因组DNA为模板,通过PCR扩增技术得到16 kDa启动子片段,序列分析结果表明:获得的启动子片段的大小为931 bp,与已报道的该启动子序列相比较,其核苷酸序列同源性为99.9%.该启动子区域含有TATA-box,CAAT-box,GCN4基序,Prolamin-box等胚乳特异表达启动子所必需的正调控元件.利用该启动子构建了植物种子特异表达载体pC16 kDP.     

植物转化酶及其基因表达与调控

蔗糖利用依赖于蔗糖的降解.植物转化酶通过降解蔗糖为己糖,不仅为呼吸作用提供底物,而且作为信号分子调节多种基因的表达,同时转化酶基因的表达也受到多种因素的调控.综述了近年来植物转化酶的种类、特性、功能及其基因表达在转录水平和翻译水平调控机制的研究进展.     

Neuroglobin mutation associated with hypoxia adaptation in Tibet chicken

Globin protein family plays an important role in storing and transporting oxygen. As a newly reported globin, the revealed function of neuroglobin includes binding and storing oxygen as well as facilitating the utilization of oxygen in neuronal cells. In the present study, mutations in exons of chicken Ngb gene were identified with the method of sequencing and created restriction site PCR in Tibet chicken and other four lowland chicken breeds. The mutations of Lys-2224(E4)-Asn and Ser-2279(E4)-Gly were identified in exon 4 of the gene. The Lys-2224(E4)-Asn mutation existed only in Tibet chicken and the mutation frequencies increased with increasing altitude. Analysis of the haplotype and diplotype of the two mutations in Tibet chicken populations of different altitudes showed that the frequencies of TG haplotype and TTGG diplotype also increased with increasing altitude, while the reverse tendency was found on GGAA diplotype. Under the hypoxic simulation incubation, the main haplotype was TG in living embryos and GA in dead embryos. The results showed that the Lys-2224(E4)-Asn mutation may be a specific mutation associated with hypoxia adaptation in Tibet chicken.     

Neuroglobin mutation associated with hypoxia adaptation in Tibet chicken

Globin protein family plays an important role in storing and transporting oxygen. As a newly reported globin, the revealed function of neuroglobin includes binding and storing oxygen as well as facilitating the utilization of oxygen in neuronal cells. In the present study, mutations in exons of chicken Ngb gene were identified with the method of sequencing and created restriction site PCR in Tibet chicken and other four lowland chicken breeds. The mutations of Lys-2224(E4)-Asn and Ser-2279(E4)-Gly were identified in exon 4 of the gene. The Lys-2224(E4)-Asn mutation existed only in Tibet chicken and the mutation frequencies increased with increasing altitude. Analysis of the haplotype and diplotype of the two mutations in Tibet chicken populations of different altitudes showed that the frequencies of TG haplotype and TTGG diplotype also increased with increasing altitude, while the reverse tendency was found on GGAA diplotype. Under the hypoxic simulation incubation, the main haplotype was TG in living embryos and GA in dead embryos. The results showed that the Lys-2224(E4)-Asn mutation may be a specific mutation associated with hypoxia adaptation in Tibet chicken.     

Scanning for sisnatures of geosraphically restricted selection based on population genomics analysis

Natural selection, as the driving force of human evolution, has direct impact on population differentiation. However, it is still unclear to what extent the genetic differentiation has been caused by natural selection. To explore this question, we performed a genome-wide scan with single nucleotide polymorphism （SNP） data from the International HapMap Project. Single locus FsTanalysis was applied to assess the frequency difference among populations in autosomes. Based on the empirical distribution of FsT, we identified 12669 SNPs correlating to population differentiation and 1853 candidate genes subjected to geographic restricted natural selection. Further interpretation of gene ontogeny revealed 121 categories of biological process with the enrichments of candidate genes. Our results suggest that natural selection may play an important role in human population differentiation. In addition, our analysis provides new clues as well as research methods for our understanding of population differentiation and natural selection.     

Fine mapping of the dominant glandless Gene Gl2^e in Sea-island cotton （Gossypium barbadense L.）

Gle2 is a mutant gene that controls glandless trait in cotton plants and seeds. It is an important gene resource to gossypol-free cottonseed breeding. The objective of this research was to develop SSR markers tightly linked with Gle2 by using the F2 segregating population containing 1599 plants derived from the cross of G. hirsutum genetic standard line TM-1 and G. barbadense glandless mutant line Hai-1. Genetic analysis suggested that the Gle2 was an incomplete dominant gene. Based on the backbone of genetic linkage map from G. hirsutum × G. barbadense BC1 published by our laboratory,Gle2 was lo-cated between CIR362 and NAU2251b,NAU3860b,STV033,with a genetic distance 9.27 and 0.96 cM,respectively. This result is useful for cloning Gle2 gene by map-based cloning method.