实时荧光定量PCR检测PrP基因在金黄地鼠脑组织的动态表达

金黄地鼠是研究动物传染性海绵状脑病的理想模型动物之一,其脑组织内朊蛋白基因动态表达数据的测定对探讨该类疾病的发生、发展和分子致病机理具有重要意义。我们利用实时荧光定量RT-PCR技术,对不同年龄金黄地鼠大脑、小脑、丘脑和脑干PrP基因的表达进行了定量。结果发现,脑的四个检测部位都呈现高的表达量,但是同一年龄段不同组织每纳克总RNA中朊蛋白基因的表达量和每毫克组织中朊蛋白基因的表达量有显著的差别,不同组织在不同年龄出现表达高峰。本研究的结果对于探讨朊蛋白的基本功能和脑组织在传染性海绵状脑病病理发生中的作用,提供了基础数据。     

Biological characteristics of JAK2 transduced CD34+ cells from cord blood during ex vivo expansion

AIM: To explore the feasibility and biological characterization of long-term regulated expansion of JAK2 transduced human CD34⁺ cord blood cells in vitro.METHODS: A retrovirus (RV) vector which contains JAK2 catalytic domain and two binding sites for a chemical inducer,dimerization (AP20187),was cloned (designated MGI-F₂JAK2).CD34⁺cells were enriched from cord blood with a MiniMACS system.The purified CD34⁺ cells were transfected with supernatant from the retrovirus packaging cell line that expressed JAK2.Following transduction,cells were expanded into four groups: AP20187 alone,FL alone,TPO,alone,AP20187+FL+TPO,respectively.The expanded cells were monitored by GFP expression,immunophenotyping,progenitor colony assay,karyotype analysis as well as tumorigenesis in nude mice.RESULTS: The purity of selected CD34⁺ cells was over 91% and gene transfer rate was 49.32%±6.21%.Only the group of AP20187 +FL+ TPO was obtained a significant sustained outgrowth of the transduced CD34⁺ cord blood cells.The percentage of GFP+ cells consistently produced a rise to the 90% peak level by the end of 8th week of culture.Flow cytometry analysis showed that the phenotype of the expanded cells was CD33⁺,CD61⁺ and Gly-A⁺ partial positive;CD38⁺ and HLA-DR⁺ strong positive,while CD2,CD7 and CD19 were almost negative.Colony assays performed in methycelluos,which can give rise to BFU-E,CFU-GM and CFU-Mix,the CFU-GM was predominantly in all colonies.The tumor was not observed in nude mice and the karyotype analysis was normal from expanded cells.CONCLUSION: The results demonstrate that AP20187-mediated activation of JAK2 signaling is capable of stimulating expansion JAK2 transduced CB CD34⁺ cells in combination with FL and TPO.This system may have applications for studies in signaling transduction,hematopoiesis,and for gene and cell therapy.     

Nucleostemin gene expression in human breast tumor tissues and its relation with clinical pathology

AIM: To study the expression of nucleostemin (NS) gene in human breast tumor tissues and the relations of NS gene expression level with histological grades,histological types and TNM stages of the tumor.METHODS: Total RNA was isolated from human breast tumor tissue.The methods of electrophoresis and RT-PCR were used in measuring NS gene expression level,and the relations of NS gene expression level with histological grades,histological types and TNM stages of the tumor were analyzed.RESULTS: The results indicated that there was no NS gene expression detected in normal breast tissues,and NS gene expression in malignant breast tumor tissues (P<0.01) was higher than that in the benign breast tumor tissues.The higher histological grades of the breast cancer showed the stronger NS gene expression (P<0.01),the higher TNM stages of the breast cancer showed the stronger NS gene expression (P<0.01),and the level of NS gene expression had not correlation with the histological types (P>0.05).CONCLUSION: It is suggested that there is no relation of NS gene expression level with histological types of the breast cancer,but there is a marked correlation of NS gene expression level with the histological grades and TNM stages.     

家蚕产卵量的基因效应参数估计

用产卵量差异较大的两个近交系782和Ekp作亲本,配制了F1、F2正反交及F1与两亲本的回交B1和B2,加上两亲本P1和P2共8个世代,采用多世代平均值的多元回归分析方法对产卵量的基因效应参数进行了估计,得到了如下结果：（1）建立了分析家蚕产卵量的常染色体基因加性、显性和性染色体基因加性效应的非二体遗传模型,并通过实际数据检验表明产卵量遗传符合这个模式;（2）产卵量的遗传主要取决于常染色体上基因加性效应,其次是性染色体基因加性效应,显性效应很小。     

猕猴桃成熟果实中脂氧合酶基因的克隆

采用ＰＣＲ扩增方法,从成熟猕猴桃果实组织中克隆到一个８２４ｂｐ的脂氧合酶（ＬＯＸ）基因片段（ｐＫＬＣ１）,该片段含有２７５个氨基酸组成的开放阅读框架,它与拟南芥菜、黄瓜、豌豆、土豆、水稻、大豆、烟草和番茄的核苷酸同源性为５９．０％～７４．６％,氨基酸同源性为６３．１％～７６．３％。Ｓｏｕｔｈｅｒｎ杂交表明,ｐＫＬＣ１片段为一低拷贝数基因家族所编码。     

Decomposition of (1–3,1–4)-β-Glucan and Expression of the (1–3,1–4) -β-Glucanase Gene in Rice Stems during Ripening

Abstract

To elucidate the possible participation of hemicellulose decomposition in lodging resistance, we studied the change of hemicellulose and cellulose content in the stems of rice during the ripening stage by methylation analysis and the expression of related genes by Northern blotting. In the rice stem in ripening stage, content of (1-3,1-4)-β-glucan, a component of hemicellulose, decreased markedly although the content of arabinoxylan, a major component of hemicellulose, and cellulose showed little change during the same growth period. On the other hand, expression of the Gns 1 gene, which may encode (1-3,1-4) -β-glucanase that catalyzes the degradation of (1-3,14) -β-glucan, increased sharply in the stem. The mechanism of decomposition of (1-3,1-4) -β-glucan in rice stem and the possible association with lodging resistance is discussed.     

基因多态性与牛疾病相关性的研究进展

基因多态性是指基因组序列上的变异,它是决定家畜对疾病易感性与抵抗力、疾病临床表现多样性以及不同个体对药物反应性的重要因素.研究基因多态性有利于定位和鉴定致病基因和疾病易感基因,使得从遗传学的角度控制疾病成为可能,从而达到减少家畜疾病防治的成本,改良家畜品种,为人类提供健康的动物产品的目的.笔者主要从抗病育种的角度综述了近年来基因多态性与牛疾病易感性和抗性的相关研究,并结合遗传信息学和表观遗传学,阐述家畜疾病控制的新进展.