Effects of blocking the expression of PKARⅠβ gene with antisense nucleotide on Shuang long-Jie gu Pill (SLJGP) containing serum osteoblast proliferation

AIM: To further investigate the role of PKARⅠβ in the growth-promoting effects of shuang long Jiegu pill (SLJGP), a Chinese medicine, on cultured osteoblasts. METHODS: pcDNA- antiPKARⅠβ, a recombinant expressing the antisense sequence of PKARⅠβ, was constructed and transformed HFOB1.19 by lipofectin. MTT was undertaken to assess the cell growth with the treatment of high dosage of SLJGP containing serum. RESULTS: Antisense gene blocked the growth-promoting effects of SLJGP containing serum on HFOB1.19. CONCLUSION: The function of SLJGP is closely related to cAMP-dependent protein kinase A.     

浅谈工程造价的预结算审核

工程造价的预结算审核是合理确定工程造价的必要程序及重要手段.本文从工程造价中存在的问题,工程造价审核的内容、方法等三个方面论述了工程造价及其预结算的审核工作过程.     

基本建设单位对工程建设项目的管理程序

工程建设项目的管理程序是基本建设单位进行项目管理的关键.本文从项目的前期准备、工程项目建设、工程项目验收三个阶段论述了基本建设单位对工程建设项目的管理程序.     

造价管理确定方法常见问题及对策

本文剖析了工程造价管理确定方法存在的问题,并提出了相应的解决策略。以期为我国建设工程造价确定方法的改革作出一定的贡献。     

桑树花青素合成酶(ANS)基因的克隆及在2种果色桑树中的表达特征

花青素合成酶(anthocyanidin synthase,ANS)是植物花青素生物合成途径末端的关键酶,催化无色花色素到有色花色素的转变。从广东桑品种粤椹大10中克隆得到一条ANS序列,其ORF为1 077 bp,编码358个氨基酸,预测蛋白质分子质量为41 kD,等电点为5.62,具有2-酮戊二酸双加氧酶的保守结构域。多重序列比对表明物种间的ANS序列高度保守,系统进化树中桑树ANS与芍药科、芸香科、葡萄科等植物的同源序列聚在一个类群上。RT-PCR检测桑树ANS在结紫色果的粤椹大10的幼叶和桑椹中特异性表达,并且随着果色加深其表达水平呈上升趋势,而在结白色果的桑品种珍珠白的幼叶和桑椹中检测不到ANS的表达,暗示桑树ANS在桑椹的颜色形成中起到重要作用。     

抗菌肽的研究与应用

抗菌肽具有分子量小、热稳定性好、无免疫原性、抗菌谱广等特点;并且,抗菌肽对畜禽具有促生长、保健和治疗疾病等功效;具有无毒副作用、无残留、无致细菌的耐药性等特性;在动物养殖和提高畜产品品质方面,抗菌肽也具有重要的潜在应用价值。文章结合当今抗菌肽的研究现状和发展前景,对抗菌肽的分类、抗菌机理、应用状况等进行了综述。     

探析市政道路桥梁加固设计的方法

近些年,随着建筑行业的发展,我国的道路桥梁建设也得到不断发展.而随着道路桥梁建设的不断推进,越来越多的人开始关注道路桥梁的安全问题.因此对加固技术在道路桥梁施工中应用的必要性进行分析的基础上阐述了桥梁维修加固机理及存在的问题,最后全面解析了市政道路桥梁维修与加固施工技术实践要点,希望可以给相关工作人员提供帮助.     

高效诱导植物RNA沉默的反向重复构件的设计

RNA沉默(RNA silencing)是真核生物体内普遍保守的、发生在RNA水平的、基于核酸序列同源性相互作用的一种对基因表达的调控机制.通过设计反向重复构建在植物体内产生双链RNA来诱导RNA沉默的方法是一条非常有效的植物抗病毒和功能基因组研究途径.本研究通过两个病毒基因的反向重复构建发展了两种通过小型中间载体设计反向重复构建的方法.其共同特点是构建简易,操作方便,可为通过设计反向重复构建实现植物体内高效持久的RNA沉默提供方法上的参考,促进RNA沉默技术更加广泛深入地应用于植物抗病毒基因工程研究和植物功能基因组研究.     

Alteration of apoptotic susceptibility and bcl-2 gene in malignant transformation of human bronchial epithelial cells

AIM: To demonstrate the susceptibility of cell apoptosis varies during the progress of cell malignant transformation from human being in vitro. METHODS: A SV₄₀T-transfected human bronchial epithelial immortalized cell line (called M) was selected in this work, which has acquired some characteristics of malignant transformation at the later passage. The alterations of apoptosis and bcl-2, P53 genes between early and later passage of M cells were investigated by means of TDT labeling in situ, chromosome FISH, RNA and protein testing, etc. RESULTS: Incidence of apoptosis induced by cis-platin was significantly lower in later than in early passages of M. Levels of bcl-2 mRNA and protein in later passages were higher than early passages of M, and overexpression of bcl-2 was accumulated following the development of cellular malignancy. P53 protein level was as high in early as in later passages. CONCLUSION: Overexpression of bcl-2 decreases the cellular sensitivity to apoptotic inductors plays an important role during progress of carcinogenesis in human bronchial epithelial cancers. The inactivation of P53 protein in the SV₄₀-T transfected M cell line may be one of reasons of bcl-2 overexpression, but not associated with the accumulation of bcl-2 expressed level during cell transformation.     

Expression of phenylalanine ammonia-lyase cDNA from rice in E. coli BL21DE3

AIM: To study the expression and its kinetics of rice phenylalanine ammonia-lyase gene encoding into E. coli as the basis of treatment for phenylketouria. METHODS: The phenylalanine ammonia-lyase-1-cDNA(rPAL-1-cDNA) from rice was recombined into E. coli high expression vector pET-28c and transformed into E. coli host strain BL21DE3. Engineering bacteria was then inducted by isopropyl-β-D-thiogalactoside (IPTG) for 1, 3, 5, 7 hours, in order to obtain high level expression. RESULTS: After induction, the expression level of fusion protein was 21.40%, 30.60%, 35.40%, 35.43% respectively. The fusion protein exhibited a band of 78.6 kD on SDS-PAGE analysis, but was not found in controls.The target protein was mainly existed in the form of inclusion body. CONCLUSION:Rice PAL gene expressing E. coli was established by gentic engineering technique.