Preparation of gfp-bcl-XL-contained recombinant adenovirus vector by the homologous recombination in bacteria

AIM: To prepare gfp-bcl-X_L-contained recombinant adenovirus(rAd-gfp-bcl-X_L).METHODS: Bcl-X_L gene was amplified from pEGFP-C₃-bcl-X_L, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-X_L. Then pAdTrack-CMV-bcl-X_L was linealinzed with PmeI and co-transformed into BJ5183 bacteria with adenovirus genomic plasmid of pAdEasy-1. The identified recombinant adenovirus plasmid was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles. The target gene was detected by PCR.RESULTS: There were about 35% positive recombinant bacterial clones after the co-transformation of pAdTrack-CMV-bcl-X_L and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-X_L into 293 cells. PCR test indicated that the recombinant Ad contained bcl-X_L gene. The titer of the purified rAd-gfp-bcl-X_L was 6.5×10¹² PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-X_L. This affords a good gene transfer vector for the gene therapy in human’s diseases.     

Effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, β-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.     

极早熟葡萄新品种‘90- 1’

 ‘90- 1’是从‘乍娜’的芽变中选出的葡萄新品种。该品种除保持母本的优质、丰产、适合保护地栽培等优良性状外, 其最突出的特点是极早熟, 果实6 月下旬成熟, 果实发育期仅35 d。     

喀喇昆仑山盖孜河水文水资源特性分析

喀喇昆仑山河流自有水文资料记载以来 ,近 10年洪灾发生频率明显高于以往 ,人民的生命财产损失程度也大幅度增加。在分析了盖孜河流域的径流及气候多年变化过程后 ,认为洪灾频发的原因是近年来流域内降水及气温呈上升趋势 ,导致高山冰川退缩 ,河流径流增加 ,洪灾增大、频率增高 ;小区域多年气候变化过程从侧面印证了我国西北地区气候由暖干向暖湿变化。     

苹果梨的育种价值

该文指出,苹果梨综合性状优良,是很好的梨育种种质资源;以苹果梨及其后代为亲本已选育出30余个新品种,大大丰富了梨种质资源;苹果梨的大果、耐贮、小果心、抗寒等优良性状在后代品种中有较强的表现;以苹果梨与多倍体品种杂交后代为多倍体的比率较高。并展望了苹果梨在今后育种中的主要研究方向。     

百合组培中鳞片处理及其颜色变化与鳞茎形成的关系

 研究了百合鳞片发育程度、切割段数、形态及生理变化、培养基的P 和K含量与鳞茎形成的关系。结果表明: 取中层鳞片、每片横切6 段的繁殖系数最高, 基部鳞片结鳞茎数最多, 且鳞茎发生最早。鳞片由基部向顶部繁殖系数递减。培养过程中鳞片颜色和形态变化依次为乳白色、浅黄、浅紫色、绿紫、鳞片干枯、鳞茎增大, 鳞片由浅黄变为绿紫色过程中叶绿素含量增加, 淀粉和可溶性糖含量下降。适当增加培养基中P、K的含量可提高鳞茎形成的数量和质量。     

敌百虫的液相色谱分析方法

本文采用即以乙腈+水(50+50)为流动相,用CN柱和紫外检测器分离测定敌百虫原药及40%敌·毒乳油中敌百虫反相高效液相色谱法。结果表明,本方法的标准偏差为0.3233,变异系数为0.3551%,平均回收率为99.65%,线性相关系数为0.9997。     

微卫星DNA在动物亲子鉴定中的应用及研究进展

本文概述了微卫星DNA进行亲子鉴定的基本原理与方法,将其用于亲子鉴定的优点以及研究进展,最后对未来的应用进行了展望。     

A mutation confers Monochoria vaginalis resistance to sulfonylureas that target acetolactate synthase

Acetolactate synthase (ALS) is the target enzyme for four distinct families of compounds: sulfonylureas (SUs), imidazolinones, triazolopyrimidine sulfonanilides, and pyrimidinyl oxybenzoates. We cloned and sequenced the fragments encoding ALS genes from biotypes of Monochoria vaginalis susceptible (S) and resistant (R) to SU-herbicides. The nucleotide sequences of the 39 bp Domain A region for R M. vaginalis biotype differed from that of the S biotype by a single nucleotide substitution at variable Pro codon of Domain A (CCT to TCT), predicting a Pro in the S but a Ser in the R biotype. No nucleotide differences between S and R M. vaginalis were observed in Domain D. We suggest that the amino acid substitution at Domain A region is responsible for resistance to SU-herbicides in M. vaginalis collected from Ushiku City, Ibaraki Prefecture, Japan.     

野榆钱菠菜三种异型种子幼苗的生长特性

野榆钱菠菜是新疆重要的荒漠野生牧草资源,具有褐色(非休眠类型)、黑色大、黑色小(休眠类型)3种异型种子。以种子异型性植物野榆钱菠菜为材料,对3种异型种子在自然条件下进行培育,对比观察研究3种异型种子的萌发物候及幼苗的生长特性。结果表明:1) 异型种子萌发物候不同步,萌发策略不同,褐色种子集中萌发,出苗率为74.30%,萌发期为3 d;两种黑色种子萌发期长,分散萌发,出苗率分别为6.58%(黑色大种子)和1.55%(黑色小种子),萌发期分别为21和17 d。2) 异型种子子叶存留时间长,但存留时间有差异,种子越小子叶存留时间越短。3) 褐色种子幼苗在初期其子叶长宽、根长、地上地下生物量均显著大于黑色种子幼苗,但随着幼苗的生长,3种异型种子幼苗在这些指标方面的差异逐渐缩小。4) 异型种子幼苗具有不同的生物量分配策略,褐色种子幼苗主要将养分输送到地上部分,两种黑色种子幼苗主要将养分输送到地下部分,这种不同的生物量分配策略能够使种子异型性植物在面对荒漠异质环境时具有不同的应对方式,是种子异型性植物能够长期适应荒漠异质环境的重要生态适应机制。研究结果为揭示种子异型性植物长期适应荒漠异质环境提供新的科学依据,也为“两头下注”对策理论提供新的理论基础。