Cloning of aminopeptidase N promoter and its activity in hematopoietic cell and different tumor cell lines

Aminopeptidase N (APN) promoter region was cloned and sequenced from peripheral blood mononuclear cells. The recombinant reporter construct containing the promoter and luciferase gene, designated pXP1-APNLuc, was introduced into myeloblastic cell line, T lymphocyte cell line and various tumor cell lines. Luciferase assay showed that APN upstream promoter is myeloid-specific for high expression in myeloblastic cell line and much lower expression in T lymphocyte cell line. The promoter activity was relatively high in lung adenoma cell line compared with other tumor cell lines including hepatoma cell line, tong cancer cell line and esophageal cancer cell line in which the promoter activity significantly diminished or was almost undetectable. The characteristics of APN promoter may provide a new strategy for specific myeloprotection while tumor patients are being treated with chemotherapy and/or radiotherapy.     

Introduction of rice chitinase gene into wheat via low energy Ar+ beam implantation

A chitinase gene (RCH8) in plasmid vector pCAMBIA1308 was delivered into 3 wheat cultivars (Yangmai 158, Wan 9210, Wanmai 32) by low energy Ar⁺ beam-mediated method. Preliminary calli from treated mature embryos were first selected on hygromycin (Hm, 20 or 30 mg/L) containing medium. After the resistant calli formed, they were transferred to the regeneration medium with 10 or 20 mg/L Hm. All the three wheat varieties obtained transgenic plants. PCR and PCR-Southern assays showed that most plants regenerated from the resistant calli were positive transgenic plants. Southern blot of the positive green plants confirmed stable integration of alien DNA into wheat genome. The plant transformation frequencies varied with the variety and ion dose implanted. Wanmai 32 possessed the highest transformation frequency, reaching 3.8% at a suitable implantation dose. The transformation frequency of Yangmai 158 and Wan 9210 varied from 0.5% to 2.5% and from 0.5% to 1.4%, respectively. Progeny test for resistance to wheat scab showed that the leaf extract of R₁ generation inhibited the growth of wheat scab strain R₀ and F₁₅.     

转基因植物的生态影响

自从1983年转基因植物诞生以来,至今各种类型的转基因植物进入大田实验已不计其数,很多产物已商品化。作为生物界的新品种,转基因植物在带来巨大的经济效益和社会效益的同时,也带来了潜在的风险。从生态学角度论述了转基因植物所带来的影响及采取相应的措施来加以调控和管理。     

β葡聚糖酶杂合基因bglHAM的克隆及在P.pastoris中的表达

采用PCR的方法,以Bacillus macerans总DNA为模板,构建出β-1,3-1,4葡聚糖酶杂合基因bglHAM,与巴斯德毕赤酵母表达载体pPIC9K重组,得到重组质粒pPIC9K-HAM,电脉冲转化法转化酵母菌GS115,KM71,在MM,MD平板上筛选表型,YPD-G418平板上筛选多拷贝重组子,重组菌株经甲醇诱导后,刚果红平板染色法检测到β-葡聚糖酶活性,SDS-PAGE证明表达产物的分子量约为24kD,摇瓶诱导培养筛选到的重组菌株,胞外每mL发酵液最高酶活达240U。     

大鼠肝ADAMs种类鉴定及肝再生过程中ADAMs mRNA差异分析

以 2 / 3肝切除 (patialhepatectomy ,PH)大鼠为材料 ,利用PT -PCR技术 ,通过ADAMs通用引物初步分析了肝脏中ADAMs种类及PH后恢复 4h和 36h时ADAMsmRNA差异。结果表明 :PH后不同恢复时间ADAMsmRNA的扩增谱带没有差异 ,但扩增谱带的强弱有变化 ;cDNA克隆和测序分析表明 ,大鼠肝脏有ADAM15和ADAM 19mRNA同源序列。     

λ噬菌体的溶原和溶菌过程与基因表达的时序控制

本文从λ噬菌体的生活周期入手,介绍了λ噬菌体的基因组结构及基因的表达过程和方式,展示了λ基因组表达的时间流程．     

利用病毒研究植物的生物学功能

现代病毒分子生物学的研究为植物的生物学功能研究提供了有效的工具。这类研究使我们对植物的细胞间通讯、抗病毒和基因表达调控等机制有了更深入的了解。也为进一步的研究提出了许多新的课题。     

植物细胞壁中富含甘氨酸的蛋白质

在植物的某些组织中存在富含甘氨酸的蛋白质,根据ＧＲＰｓ中甘氨酸的含量和氨基酸的种类、数目及排列顺序的差异,推测植物的ＧＲＰｓ存在３种类型,这些ＧＲＰｓ在植物的生长发育及植物防御过程中起重要作用,已有大量的ＧＲＰ基因被分离鉴定,并对一些ＧＲＰ基因的启动子进行了分析,ＧＲＰｓ基因的表达受植物发育的调节,具有组织或器官特异性,很多ＧＲＰ基因的表达还受环境因素的影响。     

Construction of the glucose isomerase deficient strain ofStreptomyces M1033 by homologous recombination

After the establishment of the transformation conditions ofStreptomyces diastaticus No.7 Strain M1033, the integration plasmid pXW for homologous recombination, which contains a 600 bp fragment of incompleteGI (G138P. G247D) gene, has been constructed in order to realize the stable overexpression of theGI (G138P. G247D) which is valuable for large-scale industrial production. The Gigene’s disruption has been realized by pXW’s integration into M1033 chromosomes via homologous recombination andGI deficient strain ofStreptomyces M1033 has been obtained. The reliability of introduction of mutation has been proved by analysis of recombinant fragment and affirmance of existence of the mutation, as well as detection of the stability of the deficient strain.