Cloning and over-expression of an alkaline protease from Bacillus licheniformis

The alkaline protease gene, apr, from Bacillus licheniformis 2709 was cloned into a Bacillus shuttle expression vector, pHL, to yield the recombinant plasmid pHL-apr. The pHL-apr was expressed in Bacillus subtilis WB600, yielding a high expression strain BW-016. The amount of alkaline protease produced in the recombinant increased by 65% relative to the original strain. SDS-PAGE analysis indicated a Mr of 30.5 kDa. The amino acid sequence deduced from the DNA sequence analysis revealed a 98% identity to that of Bacillus licheniformis 6816.     

Unassisted refolding of urea-denatured arginine kinase from shrimp Feneropenaeus chinensis: evidence for two equilibrium intermediates in the refolding pathway

The refolding process and the equilibrium intermediates of urea-denatured arginine kinase (AK) were investigated by 1-anilino-8-naphthalenesulfonate (ANS) intrinsic fluorescence, far-UV circular dichroism (CD), size-exclusion chromatography (SEC), and enzymatic activity. In dilute denaturant, two equilibrium refolding intermediates (I and N') were discovered, and a refolding scheme of urea-denatured AK was proposed. During the refolding of urea-denatured AK, the fluorescence intensity increased remarkably, accompanied by a significant blue shift of the emission maximum and a pronounced increase in molar ellipticity of CD at 222 nm. The first folding intermediate (I) was inactive in urea solution ranging between 2.4 and 3.0 M. The second (N') existed between a 0.4- and 0.8-M urea solution, with slightly increased activity. Neither the blue shift emission maximum nor the molar ellipticity of CD at 222 nm showed significant changes in these two regions. The two intermediates were characterized by monitoring the ANS binding ability in various residual urea solutions, and two peaks of the emission intensity were observed in urea solutions of 0.6 and 2.8 M, respectively. The SEC results indicated that a distribution coefficient (K(D)) platform existed in urea solutions ranging between 2.4 and 3.0 M urea, suggesting that there was a similarly apparent protein profile and size in the urea solution region. The refolding kinetics showed that the urea-denatured AK was in two-phase refolding. Proline isomerization occurred in the unfolding process of AK, which blocked the slow phase of refolding. These results suggested that the refolding process of urea-denatured AK contained at the least two equilibrium refolding intermediates.     

中国野生鸭茅种质资源锈病抗性研究

鸭茅是一种著名的温带牧草,容易感染锈病。本研究从植株感病率、感病严重度、病情指数和反应指数4方面对国内37份鸭茅种质资源(野生资源35份)进行锈病的病情调查及抗病性研究。研究初步揭示了鸭茅锈病的发病规律,研究结果显示02-106、02-107、90-130、02-115等4份材料对鸭茅锈病具有较好抗性,可作为育种材料进一步深入研究。     

经脐单孔腹腔镜胆囊切除术的临床研究

目的:评价经脐单孔腹腔镜胆囊切除术在临床上应用的安全性及可行性.方法;回顾性分析2008年10月到2009年8月间我院收治的6例经脐单一切口腹腔镜胆囊切除手术患者的临床资料,并与同期15例传统腹腔镜胆囊切除术患者进行比较.结果:所有腔镜手术均成功,无术后并发症,恢复较好.经脐手术组的平均住院时间为7.33±0.82,与传统腔镜组的平均住院时间(6.73±0.96)无明显差异(P＞0.05);两者的平均住院费用也无差异(P＞0.05).结论:在慎重选择病例的基础上,经脐单孔腔镜胆囊切除术是一种安全、可行的手术.     

青少年双生子体型指征相关指标的遗传度研究(英文)

目的:用经典的双生子设计探讨遗传因素对青少年身高、体重、体质指数(BMI)等体型指征的影响程度。方法:从青岛市双生子库中选择自愿参加本研究的362对11-19岁双生子,测量身高、体重。同性别的双生子通过16个多态标记进行卵型鉴定,在此基础上,应用Mx软件构建结构方程模型分析计算遗传度。结果:对362对有效双生子数据进行分析,其中同卵双生子194对,异卵双生子168对,身高的最佳拟合模型ACE模型,体重和BMI的最佳拟合模型为AE模型。调整年龄性别后,身高的遗传度为66%,体重的遗传度为84%,体质指数的遗传度为75%。结论:在青少年体型指征的相关指标中,身高、体重、BMI受遗传因素影响都较大。     

北江间(鱼骨)的繁殖生物学

间(鱼骨)(Hemibarbus medius Yue,1995)属鲤形目,鲤科,(鱼句)亚科,(鱼骨)属,为1995年报到的一个新种[1].在此之前,学者一直将其归为唇(鱼骨)[2-5].间(鱼骨)广泛分布于华南及西南部分地区,如珠江、南盘江、乌江和万泉河等水系[6].其肉质鲜嫩、营养价值高、产量较大,一直为产区的重要经济鱼类.     

连栽桉树人工林生产力和植物多样性及其相关性分析

对不同连栽代次的尾巨桉人工林进行长期定位观测,研究林分各组成部分生物量及林下植物多样性的变化,并采用趋势面分析方法分析生物量与植物多样性之间的关系.结果表明:(1)第1、2、3代7年生尾巨桉人工林的群落生物量基本相同,分别为102.22、105.11、102.66 t/hm2,其中乔木层林分生物量随着连栽代次呈增加趋势,第1代到第3代林分分别为81.48、88.36、92.28 t/hm2,平均净生产力分别为11.64、12.62、13.18 t/hm2·a,林分各器官生物量分配无明显差异,但在各生长年份,随着代数增加也呈增加趋势.(2)尾巨桉林下植物及枯落物生物量明显下降,第1、2、3代灌木层生物量分别为8.47、6.89、3.09 t/hm2,草木层生物量分别为1.93、1.21、0.35 t/hm2,枯落层生物量分别为10.34、8.64、6.94 t/hm2.(3)连栽尾巨桉人工林植物多样性指数、均匀度指数、丰富度指数随着连栽代次的增加而明显下降,即林下植物多样性随连栽代次的增加而减少,物种生态优势度增加.(4)采用趋势面分析方法结果表明,不同连栽代次尾巨桉林地植物多样性增加则林地根系生物量和林下植物生物量显著增加,而对林分总生物量及其它器官生物量影响不大.     

初次卵裂时间是猪克隆胚胎发育潜能的重要标识

初次卵裂时间与哺乳动物胚胎发育潜能有关．比较了不同初次卵裂时间(20～24 h,早期;25～36 h,中期;37～48 h,晚期;20～48 h,对照)的猪孤雌(parthenogenetic,PA)、体细胞核移植(somatic cell nuclear transfer,SCNT)胚胎的囊胚发育率、扩张囊胚发育率和囊胚细胞数,评价其体外发育能力．发现早期卵裂的PA胚胎发育到第6天的囊胚发育率显著高于中期、晚期以及对照组(P < 0.05;54.0% vs. 19.6%,5.4%,18.7%)．扩张囊胚发育率,早裂胚胎同样优于其他组．早期卵裂的SCNT胚胎发育到第6天的囊胚比率高于中期卵裂胚胎(32.2% vs. 23.5%),而晚期卵裂胚胎发育到囊胚的比率最低(6.3%)．早期卵裂的SCNT胚胎发育到第6天的扩张囊胚比率显著高于其余各组 (P < 0.05;18.9% vs. 5.9%、3.1%、7.4%)．囊胚细胞数在早期、中期、晚期三组之间表现出下降趋势．将早期卵裂的SCNT胚胎与未经挑选的对照组胚胎分别进行移植,观察其体内发育能力．移植早裂SCNT胚胎的受体在产仔数和克隆效率上均明显高于未经挑选胚胎的受体(4.7 vs. 2.1;3.9% vs. 0.9%),说明早裂胚胎着床后具有更强的发育能力．以上结果表明：初次卵裂时间可以作为猪克隆胚胎发育潜能的重要标识,选择早裂的胚胎进行移植,有助于提高克隆效率．     

不明原因男性不育人群中睾丸特异性乳酸脱氢酶基因突变的发现及其意义

为了研究睾丸特异性乳酸脱氢酶,即乳酸脱氢酶C4(LDH-C4)基因突变在男性不育发病中的作用,利用LDH-C4特异性底物对100名不明原因男性不育症患者的精子LDH-C4进行活性显色,用变性高效液相色谱(DHPLC)技术对LDH-C4活性低下的患者进行LDHC基因PCR产物的突变筛查,对DHPLC峰形异常的PCR产物进行序列测定.筛选到一组精子LDH-C4活性明显下降的患者,其中1名患者的LDHC基因PCR产物在DHPLC中呈异常洗脱峰.对这一PCR产物进行序列测定,发现患者LDHC基因第5外显子的115位碱基发生了T→A的杂合改变(GenBank登录号GU479375),该突变使LDHC基因的178位密码子由原来的TTG(编码亮氨酸)变为TAG(终止密码子),形成截短的C亚基.T克隆-测序进一步证实了该无义突变的杂合状态.这是在人类LDHC基因上发现的第一个突变,提示LDHC基因突变可能是男性不育发病的原因之一.