农杆菌介导油菜油体基因与碱性成纤维细胞生长因子融合基因转化油菜研究

1材料与方法 1．1材料 1．1．1供试材料。白菜型油菜苏州青,市售。 1．1．2菌株及质粒。采用根癌农杆菌LBA4404,该菌株含有植物双元表达载体pCAMBIA1390,油菜油体基因与bFGF的融合基因已整合到该载体上,该融合基因由油菜油体基因启动子启动。植物基因组提取试剂盒购自北京百泰克生物技术有限公司,Southern检测试剂盒为DIG High Prime DNA Labeling and Detection Starter Kit I,购于Roche公司。     

转基因技术在畜牧兽医中的发展前景

通过简要叙述转基因动物研究的发展简史、研究现状和存在问题 ,对转基因动物育种、药物生物反应器、组织器官移植 ,以及基因治疗和免疫等前景进行了综述。在此基础上 ,从畜牧兽医角度出发提出了追踪发展趋势的几点思考。     

生物反应器培养BHK-21悬浮细胞增殖伪狂犬病病毒工艺优化

旨在筛选伪狂犬病病毒（PRV）敏感的BHK-21细胞并分析其生长和病毒增殖特性,优化反应器中BHK-21悬浮细胞的培养和病毒增殖条件,建立生物反应器培养BHK-21悬浮细胞增殖PRV工艺。本研究利用响应面和单因素优化法,以细胞生长动力学特性、TCID₅₀病毒滴度等参数为指标,优化1.2 L生物反应器中BHK-21悬浮细胞的最佳培养和增殖病毒条件,在5 L生物反应器中进一步批培养验证。结果显示,筛选获得PRV高敏感的BHK-21-02贴壁细胞和BHK-21-XF02悬浮细胞各1株,BHK-21-XF02悬浮细胞在含3%血清的SLM-BHK低血清培养基和SFM-BHK无血清培养基中均能实现良好的生长和病毒增殖。利用响应面法优化得到1.2 L反应器最佳培养条件为接种密度1.20×10⁶cells·mL^-1、搅拌转速120 r·min^-1、DO值40%,5 L反应器批培养72 h细胞密度可达（7.61±0.18）×10⁶ cells·mL^-1、细胞活率为（96.93±1.18）%。利用单因素法优化得到1.2 L反应器最佳病毒增殖条件为MOI 0.001、培养温度37℃、细胞密度2.0×10⁶cells·mL^-1、搅拌转速80 r·min^-1,5 L反应器批培养接毒后48 h病毒滴度达到最大值（7.13±0.11） lgTCID₅₀·mL^-1。本研究可为PRV疫苗相关研究和规模化生产提供参考。     

转基因动物研究进展

转基因动物是现代生物技术中一个极其重要的研究领域，目前已经有转基因小鼠、兔、绵羊、山羊、猪、牛、鸡和鱼等多种转基因动物问世。本文综述了转基因动物的制作方法、转基因动物的应用研究以及所取得的重要成就，并指出了转基因动物存在的主要问题，展望了其发展前景。     

基因打靶备乳腺生物反应器研究进展

基因打靶技术是建立在胚胎干细胞和同源重组技术之上,可对基因组进行定点修饰的实验方法。用基因打靶技术制备乳腺生物反应器,克服了显微注射法的众多缺陷。本文简述了制备乳腺生物反应器过程中基因打靶的策略、靶细胞、打靶位点及国内外研究进展和展望等。     

非正规垃圾填埋场治理修复技术比选——以海南省某非正规垃圾填埋场为例

以海南省某非正规垃圾填埋场为例,对目前较成熟的非正规垃圾填埋场修复技术——就地封场、整体搬迁、垃圾减量搬迁开展了比选,基于各修复技术的特点,结合本垃圾填埋场情况,通过技术选择流程,确定了海南省某填埋场治理方案为整体搬迁。     

A Review on Medical Waste Management and Treatment in Chongqing Three Gorges Region

Medical waste ,belonging to one of top-ronking toxic wastes, should be safely deposed of. The characteristics and harmness of the medical wastes are introduced, and the practical status of medical wastes in Chongqing three gorges region, including waste management and disposal methods, is investigated. Because of the special characteristics of the waste, the paper proposes some measures for managing and separate collection and treatment in the region, including separate collection, sealed transportation, pretreatment, incineration and safe landfill, so that the ecological environment and water resource can be protected from pollution in the Three Gorges region.     

生物反应器中PRRSV TJM株在Marc-145细胞上最佳增殖条件研究

为了在Marc-145细胞上获得更高滴度的猪繁殖与呼吸综合征病毒(PRRSV)TJM株,对细胞培养条件、细胞接种量、微载体的用量以及病毒培养时间等条件进行了优化。结果表明,利用生物反应器悬浮培养Marc-145细胞在血清为金源康且含量为10%、培养基为DMEM、细胞接种密度为20~30细胞/球、微载体为5 g等条件下生长状态最好;病毒最佳培养时间为27~36 h,病毒增殖效果好且能够达到最高的病毒滴度。本试验为微载体培养条件下大规模生产PRRSV-TJM株疫苗奠定了一定理论基础。     

Treatment of Wastewater with High Concentration of Ammonia Nitrogen through ASBR Combining with Pulsed SBR

In order to analyze the nitrogen removal performance of anaerobic-aerobic process treating landfill leachate, a system combining ASBR with pulsed SBR was applied to treat real landfill leachate with high concentration of ammonia nitrogen. The HRT of ASBR was 2 d; middle water tank regulated the COD/NH +4-N (3～5) and NH +4-N concentration of pulsed SBRs influent; the cycle of pulsed SBR consisted of four anoxic stages and three oxic stages in which three equal feeds model was applied. In the fourth anoxic stage (An4), internal carbon source was used to implement denitrification without any extra carbon source addition. The results indicate that efficient nitrogen removal performance of this system was achieved in the series running period (157 d). Influent chemical oxygen demand (COD) concentration of ASBR was from 7 338 mg·L -1 to 1 0445 mg·L -1 of which above 83% was removed; pulsed SBRs influent NH +4-N concentration could be divided into four phases in which improving from 296.1±14.5 mg·L -1 to 912.0±41.7 mg·L -1, and the effluent total nitrogen (TN) concentration of pulsed SBR was below 40 mg·L -1 all the time, which means TN removal rate reached over 90%. To sum up, the COD and TN removal rate of the combined system were above 87% and 97%, respectively. Endogenous denitrification rate(DNR) of An4 changed from the fast one to the slow one, and the mean value of theoretical denitrification rate (TDNRm) of pulsed SBRs An4 could reach 1.531 mg N·h -1·gMLVSS -1. Advanced nitrogen removal of the system was realized without extra carbon source addition and physicochemical pre-treatment.