Cloning, expression, and refolding of a secretory protein ESAT-6 of Mycobacterium tuberculosis

A DNA encoding the 6-kDa early secretory antigenic target (ESAT-6) of Mycobacterium tuberculosis was inserted into a bacterial expression vector of pQE30 resulting in a 6x His-esat-6 fusion gene construction. This plasmid was transformed into Escherichia coli strain M15 and effectively expressed. The expressed fusion protein was found almost entirely in the insoluble form (inclusion bodies) in cell lysate. The inclusion bodies were solubilized with 8M urea or 6M guanidine-hydrochloride at pH 7.4, and the recombinant protein was purified by Ni-NTA column. The purified fusion protein was refolded by dialysis with a gradient of decreasing concentration of urea or guanidine hydrochloride or by the size exclusion protein refolding system. The yield of refolded protein obtained from urea dialysis was 20 times higher than that from guanidine-hydrochloride. Sixty-six percent of recombinant ESAT-6 was successfully refolded as monomer protein by urea gradient dialysis, while 69% of recombinant ESAT-6 was successfully refolded as monomer protein by using Sephadex G-200 size exclusion column. These results indicate that urea is more suitable than guanidine-hydrochloride in extracting and refolding the protein. Between the urea gradient dialysis and the size exclusion protein refolding system, the yield of the monomer protein was almost the same, but the size exclusion protein refolding system needs less time and reagents.     

食品科学与工程专业生物化学教学体会——从应用型人才培养角度出发

生物化学是生命科学、食品科学、医学等多个领域相关专业的基础课。笔者从食品科学与工程专业应用型人才培养的角度出发,对人才培养目标、教学内容、教学方法与手段和教师的"爱与激情"等几个方面进行了探讨,以期更好地指导、促进生物化学的教学工作。     

江西乐平、瑞金早二叠世栖霞组的Cystomichelinia和Protomichelinia

本文简述了江西乐平、瑞金早二叠世栖霞组Cystomichelinia和Protomichelinia的性质,与国内外有关的种进行了比较;通过分析这两属的内部构造说明它们之间的关系;根据珊瑚体的壁刺发育与否、床板生长密度、体径大小及体壁厚度等特征,拟立了该两属的种的鉴别表;描述了7种,其中4新种和1新亚种。     

我国医疗技术管理中引入医疗技术评估的路径探讨

2015年以来,在简政放权的大环境下,我国医疗技术管理面临着从重审批的分级分类管理向重监管转型的挑战和机遇,亟需探索建立更加科学合理的医疗技术管理体系。使用医疗技术评估的科学工具和手段是在医疗技术管理领域国际通行的理念和做法。本文梳理了我国医疗技术管理政策沿革,介绍了国际典型代表英国医疗技术管理的过程和方法,并为完善我国医疗技术管理提供了几条政策建议。     

西藏南部定日、聂拉木地区早-中侏罗世有孔虫

记述早、中侏罗世有孔虫13属17种,标本采自西藏南部定日县的卧龙剖面及聂拉木县中尼公路5264km路标处剖面。早侏罗世普普嘎组以产大有孔虫为特征,在卧龙剖面仅有下侏罗统的中、上部,缺失下部地层;中侏罗世聂聂雄拉组以产小有孔虫为特征,基本没有大有孔虫。     

益生菌联合早期肠内营养干预对 重症脑卒中患者营养状态及肠道菌群的影响

目的探讨益生菌联合早期肠内营养干预对重症脑卒中患者营养状态及肠道菌群的影响。方法选取2014年1月至2018年6月于我院住院治疗的重症脑卒中患者70例,随机分为观察组和对照组各35例。两组患者均予控制颅内压、营养神经细胞、抗生素预防感染、保护胃黏膜和维持水电解质酸碱平衡等基础治疗。对照组患者留置鼻饲管行常规肠内营养治疗,观察组患者在对照组治疗基础上加用双歧杆菌四联活菌片水化后鼻饲管注入,1.5 g/次,3次/d,连用10 d。观察两组患者治疗前和治疗10 d后营养状态指标[血清总蛋白(TP)、转铁蛋白(TF)、上臂三头肌肌围(MAMC)]的变化,并比较治疗后肠道菌群(双歧杆菌、乳杆菌、拟杆菌、肠球菌、大肠埃希菌)数量变化。结果治疗10 d后,两组患者血清TP、TF水平和MAMC较治疗前显著下降(P0.05),且治疗后观察组患者TP、TF水平和MAMC高于对照组(P0.05);同时治疗后观察组患者粪便中双歧杆菌、乳杆菌和拟杆菌数量高于对照组,肠球菌和大肠埃希菌数量低于对照组(P0.05)。结论益生菌联合早期肠内营养干预不仅可显著减缓重症脑卒中患者的营养状态恶化;而且能有效地纠正肠道菌群失调,维持肠道微生态的稳定。     

编写《微生物学教程》有感

《微生物学教程》获"首届全国教材奖"(二等奖)的消息后,为响应学院领导提出的总结、交流等要求,特撰写了本文.作者以"愿终生能炼成一个称职的微生物学教师"为指导思想,历经半个多世纪的努力,在教书育人的同时,编写了较多的文章和书籍,其中的代表作当属《微生物学教程》.该书至今已出了 4版,被300余所高等院校选作教材,总印数...     

百色六怀山旧石器遗址发掘简报

百色六怀山遗址位于右江右岸第四级基座阶地。2005年10—11月对该遗址进行抢救性发掘,揭露面积1000m^2,出土石制品136件,砾石37件。石制品类型包括石器、石核、石片、断块和砾石等。古人类选择阶地底部河卵石为原料进行剥片和加工石器;石器以大型为主,手镐和砍砸器是主要类型;古人类多在砾石一端采用锤击法打制石器。石器面貌具有中国南方旧石器主工业的鲜明特点。地貌和地层对比显示该遗址形成于中更新世早期。     

记内蒙古宁城道虎沟上侏罗统/下白垩统一具有长肢的蜥蜴

道虎沟化石层分布于内蒙古宁城县道虎沟以及毗邻的河北北部和辽宁西部等地,已报道4种有尾类、1个无尾类蝌蚪、1个幼年蜥蜴、3种带原始羽毛的兽脚类恐龙、2种翼龙、3种哺乳动物以及双壳类、叶肢介、昆虫和植物等化石(有学者称之为"前热河生物群")。该层位比热河群义县组低,但地层划分存在争议:中侏罗统九龙山组、上侏罗统道虎沟组和下白垩统热河群均有报道。本文研究的道虎沟蜥蜴是该地点发现的第2个蜥蜴化石,其身体细长,为一幼年个体;前颌骨、顶骨和额骨均成对;上颌骨的面突高;后额骨和后眶骨不愈合;后额骨较大,但可能未进入上颞孔;后眶骨具宽的后突;牙齿小而尖锐,结构简单且可能为侧生;头骨的眶前区较长;具27个荐前椎;虽然未成年但个体较大;未见真皮骨板;手掌和四肢较长。上述特征组合区别于所有现生蜥蜴类,四肢比例也与其他蜥蜴有较大区别。欧美中侏罗世-早白垩世的小盗蜥(Parviraptor)和蒙古早白垩世的一个幼年个体(可能属于壁虎型类)与道虎沟标本比较相似。头骨特征和较大的荐前椎数目显示道虎沟标本可能属于硬舌蜥类(scleroglossan)。但因标本为幼年个体,本文暂将它归入有鳞类属种未定(Squamata gen.et sp.indet.)。测量数据显示,道虎沟标本的手掌和前肢相对较长;该特征常见于适应攀爬的蜥蜴种类,似可显示攀爬习性。但肢体比例可能会随身体增长而发生变化,所以此生活习性并非定论。     

Control of Liriomyza langei on chrysanthemum by Diglyphus isaea produced with a standard or modified parasitoid rearing technique

Abstract: Overproduction of male parasitoids during mass rearing will increase costs for biological control because wasp shipments contain fewer females and only females kill hosts directly. We have developed a rearing technique capable of significantly reducing male‐biased sex ratios in Diglyphus isaea (Walker) (Hym., Eulophidae), a commercially reared parasitoid of agromyzid leafminers. In this study, we examined the effect of rearing technique on the efficacy of D. isaea for biological control of Liriomyza langei Frick (Dip., Agromyzidae) on chrysanthemum, Dendranthema grandiflora Tzvelev var. ‘Miramar’. We produced D. isaea on mixtures of small and large hosts (our modified technique) or on only large hosts (simulating commercial mass‐rearing) and compared: (1) control of L. langei with D. isaea produced by the two rearing techniques, and (2) damage and yield of unprotected and protected plants. The two rearing techniques produced similar numbers of waSPS per rearing cohort, but the ‘modified’ technique reduced the proportion of males by approximately 13%. The two techniques also produced females of similar size, but the ‘modified’ technique produced smaller males. In greenhouse trials simulating leafminer infestations of potted chrysanthemums during commercial production, we found no significant differences between the levels of control obtained by releasing identical numbers and sex ratios of adult waSPS produced by either rearing technique. Mine counts on plants protected by waSPS of either rearing history were similar and around 30–70% less than unprotected plants during most of the 11‐week crop cycle. At crop harvest, more than half of the foliage on protected plants was undamaged compared with <10% on unprotected crops. Damage to the flower stems of protected plants was relatively light in the top half of the canopy compared with the bottom half. Protected plants were around 10–15% taller and produced twice as many flower buds compared with unprotected plants. Our ‘modified’ rearing technique can reduce overproduction of males in D. isaea with no compromise in biological control efficacy. Adoption of our rearing technique by commercial insectaries could reduce implementation costs for not only D. isaea but also other parasitoids that show host‐size‐dependent sex allocation.