电针介导的基因转移

基因转移是实现基因治疗的关键技术之一 ,目前尚缺少简便、易行、有效、安全的方法 .首次将我国传统的针刺技术与现代转基因技术结合起来 ,创建了一种电针转基因的方法 .应用针灸针携带外源基因 ,经皮针刺 ,进行直流电刺激 ,可实现有效的基因转移 .     

Establishment of Peristenus digoneutis and P. relictus (Hymenoptera: Braconidae) in California for the control of Lygus spp. (Heteroptera: Miridae)

Lygus hesperus Knight is native to the western United States and is a perennial pest of numerous crops in California. It is responsible for triggering the early season application of insecticides on cotton, Gossypium hirsutum L., and strawberries, Fragaria L. Despite several surveys conducted in alfalfa (Medicago sativa L.) grown in central California, nymphal parasitoids associated with L. hesperus and L. elisus have not been found. Two exotic parasitoids were released into California beginning in 1998. Peristenus relictus (Ruhte), formerly P. stygicus Loan, and P. digoneutis Loan were collected from several locations in southern Europe and released at up to six locations over a 6-year period. At the original release site in Sacramento, a 0.25-ha plot of alfalfa, parasitism by P. digoneutis and P. relictus combined increased from zero to 90%, 3 years after the last releases were made. Parasitoids have been recovered from vacant fields of weedy annuals within 2 km of this site. Recoveries at more southerly release sites in central California have been poor.     

DNA methyltransferase homologue TRDMT1 in Plasmodium falciparum specifically methylates endogenous aspartic acid tRNA

In eukaryotes, cytosine methylation regulates diverse biological processes such as gene expression, development and maintenance of genomic integrity. However, cytosine methylation and its functions in pathogenic apicomplexan protozoans remain enigmatic. To address this, here we investigated the presence of cytosine methylation in the nucleic acids of the protozoan Plasmodium falciparum. Interestingly, P. falciparum has TRDMT1, a conserved homologue of DNA methyltransferase DNMT2. However, we found that TRDMT1 did not methylate DNA, in vitro. We demonstrate that TRDMT1 methylates cytosine in the endogenous aspartic acid tRNA of P. falciparum. Through RNA bisulfite sequencing, we mapped the position of 5-methyl cytosine in aspartic acid tRNA and found methylation only at C38 position. P. falciparum proteome has significantly higher aspartic acid content and a higher proportion of proteins with poly aspartic acid repeats than other apicomplexan pathogenic protozoans. Proteins with such repeats are functionally important, with significant roles in host-pathogen interactions. Therefore, TRDMT1 mediated C38 methylation of aspartic acid tRNA might play a critical role by translational regulation of important proteins and modulate the pathogenicity of the malarial parasite.     

DNA Barcoding of Economically Important Mushrooms: A Case Study on Lethal Amanitas from China

Some species of the genus Amanita are economically important gourmet mushrooms, while others cause dramatic poisonings or even deaths every year in China and in many other countries. A DNA barcode is a short segment or a combination of short segments of DNA sequences that can distinguish species rapidly and accurately. To establish a standard DNA barcode for poisonous species of Amanita in China, three candidate markers, the large subunit nuclear ribosomal RNA (nLSU), the internal transcribed spacer (ITS), and the translation elongation factor 1 alpha (tef1 α) were tested using the eukaryotic general primers for their feasibility as barcodes to identify seven species of lethal fungi and two species of edible ones which can easily be confused with the lethal ones known from China. In addition, A.phalloides—a European and North American species closely related to one of the seven taxa, A.subjunquillea was also included. PCR amplification and sequencing success rate, intra and inter specific variation and rate of species identification were considered as main criteria for evaluation of the candidate DNA barcodes. Although the nLSU had high PCR and sequencing success rates (100% and 100% respectively), occasional overlapping occurred between the intra and inter specific variations. The PCR amplification and sequencing success rates of ITS were 100% and 85.7% respectively. ITS showed high sequence variation among species group and low variation within a given species. There was a relatively high PCR amplification and sequencing success rate for tef1 α (85.7% and 100% respectively), and its intra and inter specific variation was higher than that of ITS or nLSU. All three candidate markers showed hight species resolution. ITS and tef1 α had a more clearly defined barcode gap than nLSU. Our study showed that the tef1 α and nLSU can be proposed as supplementary barcodes for the genus Amanita, while ITS can be used as a primary barcode marker considering that the ITS region may become a universal barcode marker for the fungal kingdom.     

Genetic Diversity of Global Aromatic Rice Varieties

Genetic diversity of 434 rice accessions collected from 16 countries, including 345 fragrance rice varieties and 89 non fragrance rice varieties, have been analyzed. A total of 573 alleles were detected by using 77 simple sequence repeats (SSR) primer pairs covering all rice 12 chromosomes. The value of allelic polymorphism information content (PIC) ranged from 0090 to 0845, with an average of 0516 per locus; Gene diversity (GD) varied from 0091 to 0859, with an average of 0573; The mean value of major allele frequencies (MAF) was 0540, covering from 0251 to 0953. In addition, 434 rice accessions are divided into three sub populations by cluster and population structure analysis, and FST between sub populations showed a mean of -0116, ranging from -0623 to 0494; The score of genetic distance calculated by Nei′s method appeared from 0207 to 0355. Major allele frequencies within each sub population distributed from 0408 to 0746, and gene diversity level from 0354 to 0699, while PIC from 0320 to 0658. Sequencing 6 mitochondrion genes in 18 rice varieties exhibited no different in 5 genes, whereas Mit4 contains a 3 SNPs in the gene body, which acts as an important marker to understanding the relationship between Indica/Japonica differentiation and the evolution of fragrant gene. Finally, genetic diversity and mitochondrion gene sequencing would help to know about the origin of fragrant resource and benefit rice breeding.     

非酒精性脂肪肝病大鼠肝脏SOCS-3的表达与吡格列酮干预研究

目的：探讨SOCS-3在非酒精性脂肪肝病（NAFLD）发病中的作用以及吡格列酮的干预作用。方法：29只雄性SD大鼠随机分为正常对照组（8只）,高脂饮食组（21只）。饲养8周后,从高质饮食组随机抽取5只大鼠证实造模成功后,将该组余下的16只大鼠继续以高脂饲料喂养,并随机分为NAFLD对照组（8只）;吡格酮干预组（8只）,予以吡格列酮3mg·kg^-1·d^-1灌胃。16周末,处死所有大鼠,检测血糖、血胰岛素、血脂、肝脏SOCS-3mRNA和SREBP-lcmRNA表达及肝脏病理学。结果：与正常对照组相比,NAFLD组血糖、血胰岛素、血脂、肝脏脂肪变水平及肝组织SOCS-3mRNA、SREBPlCmRNA表达显著上调。吡格列酮干预组sOCS．3mRNA、SREBP-1cmRNA表达较NAFLD组下调,且血糖、血胰岛素、血脂、肝脏脂肪变水平下降。SOCS-3mRNA表达水平与胰岛素抵抗指数、SREBP．1cmRNA表达水平、肝脂肪变成显著正相关。结论：SOCS-3可能通过胰岛素抵抗及上调肝组织SREBP-lcmRNA表达参与NAFLD发病,吡格列酮能抑制肝脏SOCS-3的表达,对NAFLD有一定治疗作用。     

蜘蛛丝蛋白的模块结构性能和重组表达

蜘蛛丝是一类天然蛋白质纤维,具有独特的机械性能（高强度、高弹性和高断裂功等）和卓著的生物学特性（生物可降解性和与生物组织的相容性等）,在生物医学、材料、纺织和军事等领域都有着很高的潜在应用价值。综述了不同蜘蛛丝蛋白的模块结构特征及与其功能的关系,扼要介绍了目前利用各种基因工程方法表达重组蜘蛛丝蛋白的研究进展。     

表没食子儿茶素没食子酸酯对人结肠癌HT-29细胞G1期的阻滞作用

目的:研究表没食子儿茶素没食子酸酯(Epigallaocatechin-3-gallate,EGCG)时人结肠癌HT-29细胞增殖的影响.方法:实验分为EGCG不同浓度处理组和阴性对照组.采用MTT比色法检测EGCG(30μg/mL、40μg/mL、50μg/mL、60μg/mL、70μg/mL)对HT-29细胞的生长影响;应用流式细胞术分析EGCG对HT-29细胞周期分布的影响;免疫印迹观测EGCG对HT-29细胞p38MAPK、cyclinD1蛋白表达的影响.结果:MTT比色结果显示.不同浓度EGCG(30μg/ml、40μg/ml、50μg/ml、60μg/ml)对HT-29细胞具有明显的生长抑制作用,并呈剂量-效应依赖关系(P<0.05);流式细胞术分析显示,EGCG诱导人结肠癌细胞G1期阻滞,且随着处理时间的延长,其诱导周期阻滞的效应越明显(P<0.05);蛋白免疫印迹显示.总的p38MAPK不随处理时间和浓度的改变而改变,但是磷酸化的p38MAPK蛋白的表达随处理时间和处理浓度的增加而明显增加,而CyclinD1蛋白的表达随处理浓度的增加而明显减少.结论:EGCG诱导HT-29细胞G1期阻滞,抑制细胞增殖,可能与活化p38MAPK,下调CyclinD1蛋白表达有关.     

Quantitative and phenotypic analysis of bone marrow-derived cells in the intact and inflamed central nervous system

Bone marrow has been proposed as a possible source of cells capable of replacing injured neural cells in diseases such as Multiple Sclerosis (MS). Previous studies have reported conflicting results regarding the transformation of bone marrow cells into neural cells in vivo. This study is a detailed analysis of the fate of bone marrow derived cells (BMDC) in the CNS of C57Bl/6 mice with and without experimental autoimmune encephalomyelitis using flow cytometry to identify GFP-labeled BMDC that lacked the pan-hematopoietic marker CD45 and co-expressed neural markers polysialic acid-neural cell adhesion molecule or A2B5. A small number of BMDC displaying neural markers and lacking CD45 expression was identified within both the non-inflamed and inflamed CNS. However, the majority of BMDC exhibited a hematopoietic phenotype.Key words: bone marrow, transplantation, transdifferentiation, central nervous system, green fluorescence protein, experimental autoimmune encephalomyelitis, multiple sclerosis