我国工业生物过程工程研究进展

工业生物技术的进步离不开工业生物过程工程研究的不断深入及发展,我国作为工业发酵大国在工业生物技术由实验室向产业化转化过程中面对诸多挑战,由此而逐渐发展起来的我国工业生物过程工程发展先后经历了多个阶段,伴随着不同阶段的发展,我国的工业生物技术水平得到不断的提升。本文重点回顾了近三、四十年来我国工业生物过程工程发展的历程,包括早期由化工过程研究引入的动力学模型化研究、基于过程控制的优化理论与方法的应用、基于过程在线监测技术发展起来的参数相关性分析方法、过程多尺度理论的建立、基于现代固态发酵的新型固态发酵罐的设计及优化技术发展等。通过对生物过程工程发展历程的回顾对先进工业生物过程发展面临的技术难题及由此引出的未来发展重点方向进行了探讨。     

杉木拟赤杨混交林林分生产力及生态效应研究

 本文从林分结构、生物量、群落特征、生产力、培肥土壤、涵养水源及林内小气候等方面对不同混交模式的杉木拟赤杨混交林及其纯林进行的研究结果表明：杉木拟赤杨是具有较高生产力和协调种间关系能力的针阔混交树种。7年生3:1带状混交林蓄积量和生物量分别比杉木纯林提高7.24％和18.22％,同时混交林还表现出比杉木纯林更好的培肥土壤、涵养水源、改善林内小气候等生态功能。应用AHP法进行不同混交模式的综合选择结果表明：3:1杉木拟赤杨带状混交模式是值得南方林区大力推广的杉阔混交模式。     

二化螟对杀螟硫磷产生抗性的机理

本文就二化螟Ckilo suppressalis Walker对杀螟硫磷抗性发生的机理进行了较为全面的研究。活体增效作用研究表明,脱叶磷(DEF)和氧化胡椒基丁醚(PBO)对抗性种群可分别增效1.3和34.8倍。这表明了多功能氧化酶(MFO)活性增强,而似乎与水解酶的活性无关。对寓体解毒酶的进一步研究表明,抗性和敏感种群的非特异性酯酶活力无明显差异,但羧酸酮酶(CarE,加入10^-4mol/L的毒扁豆碱)和MFO的活力在抗性种群中有所增高。R种群幼虫CarE活力分别是s的2.42和2.92倍(以a-和β-乙酸萘酯为底物);R种群幼虫MFO的O-脱甲基作用的活力为s幼虫的1.43倍(以对硝基茴香醚为底物);R幼虫乙酰胆碱酯酶(AChE)对乙酰胆碱的活力是s的1.3倍,两者相差不大。但从米氏常数K_m)和最大反应速度(V_max)看,R幼虫是s幼虫的1.9和1.6倍;以对氧磷为抑制剂的R幼虫150是s幼虫的2倍。这些表明了R和s的AChE是不同的。研究结果还表明,二化螟对杀螟硫磷抗性的机理至少包括：(1)CarE活性的增高;(2)MFO的O-脱甲基活力的增高和(3)AChE敏感性的降低等。     

伊犁河谷北坡野果林木本植物种间关系及环境解释

伊犁河谷北坡野果林是天山山脉中一个重要的植被类型,对其种间关系进行研究将有助于揭示野果林群落的动态特征.我们于2009年在野果林的不同群落中设置了10个样地,记录了群落的物种组成以及环境因子,包括海拔、坡度和坡向,测定了土壤含水率和有机质等土壤参数.采用x2检验、Pearson相关系数和Spearman秩相关系数检验等方法,对野果林乔木层和灌木层种间关联性进行了研究.结果表明:Pearson相关检验优于x2检验,Spearman 秩相关检验比Pearson相关检验更加灵敏;乔木和灌木优势种绝大多数种对的联结关系未达到显著水平,群落结构较为松散,表明各群落可能处于演替的早期阶段;在影响物种分布的环境因子中,海拔起决定性的作用,其次是坡向、土壤含水率、有机质、全氮和pH值.     

长江中、上游铜鱼的生长特性

铜鱼是长江中上游的重要经济鱼类和天然捕捞对象,1990—1995年作者利用在长江中上游收集的1030尾标本,对铜鱼的生长特性进行了研究。其体重和体长的关系为W=0.005413L~(3.113)(r=0.9982),生长方程为L_t=60.0229[1-e~(-0.2325(t 0.6108))];W_t=3261.9137[1-e~(-0.2325(t 0.6108))]~(3.113),体重生长拐点t=4.2龄,相应的体长41.05cm、体重973.03g。按生长指标值分析,阶段生长可明显地划分为两个时期,即4龄前的生长迅速期和4龄后的生长减缓期。因此,天然渔业的捕捞规格应控制在约1000g以上的个体,目前在葛洲坝以下的中游江段主要捕捞幼鱼的现状应该改变。     

MicroRNA-26a/b and their host genes cooperate to inhibit the G1/S transition by activating the pRb protein

The functional association between intronic miRNAs and their host genes is still largely unknown. We found that three gene loci, which produced miR-26a and miR-26b, were embedded within introns of genes coding for the proteins of carboxy-terminal domain RNA polymerase II polypeptide A small phosphatase (CTDSP) family, including CTDSPL, CTDSP2 and CTDSP1. We conducted serum starvation-stimulation assays in primary fibroblasts and two-thirds partial-hepatectomies in mice, which revealed that miR-26a/b and CTDSP1/2/L were expressed concomitantly during the cell cycle process. Specifically, they were increased in quiescent cells and decreased during cell proliferation. Furthermore, both miR-26 and CTDSP family members were frequently downregulated in hepatocellular carcinoma (HCC) tissues. Gain- and loss-of-function studies showed that miR-26a/b and CTDSP1/2/L synergistically decreased the phosphorylated form of pRb (ppRb), and blocked G1/S-phase progression. Further investigation disclosed that miR-26a/b directly suppressed the expression of CDK6 and cyclin E1, which resulted in reduced phosphorylation of pRb. Moreover, c-Myc, which is often upregulated in cancer cells, diminished the expression of both miR-26 and CTDSP family members, enhanced the ppRb level and promoted the G1/S-phase transition. Our findings highlight the functional association of miR-26a/b and their host genes and provide new insight into the regulatory network of the G1/S-phase transition.     

Reproducible and sustained regulation of Gαs signalling using a metazoan opsin as an optogenetic tool

Originally developed to regulate neuronal excitability, optogenetics is increasingly also used to control other cellular processes with unprecedented spatiotemporal resolution. Optogenetic modulation of all major G-protein signalling pathways (Gq, Gi and Gs) has been achieved using variants of mammalian rod opsin. We show here that the light response driven by such rod opsin-based tools dissipates under repeated exposure, consistent with the known bleaching characteristics of this photopigment. We continue to show that replacing rod opsin with a bleach resistant opsin from Carybdea rastonii, the box jellyfish, (JellyOp) overcomes this limitation. Visible light induced high amplitude, reversible, and reproducible increases in cAMP in mammalian cells expressing JellyOp. While single flashes produced a brief cAMP spike, repeated stimulation could sustain elevated levels for 10s of minutes. JellyOp was more photosensitive than currently available optogenetic tools, responding to white light at irradiances ≥1 μW/cm(2). We conclude that JellyOp is a promising new tool for mimicking the activity of Gs-coupled G protein coupled receptors with fine spatiotemporal resolution.     

Isolation of soluble proteins from an industrial strain Streptomyces avermitilis in complex culture medium for two-dimensional gel electrophoresis

Two-dimensional gel electrophoresis (2-DE) is a core proteomic technique to study protein expression and function in living organisms. Although 2-DE has been extensively used for the investigation of bacteria, yeast, animal and plant tissue cells, the isolation of proteins from the organisms and elimination of salt, nucleotide, polysaccharide, lipids and other contaminations from the samples often limit its application. In this study, the protocol for protein isolation from cells of Streptomyces avermitilis cultivated in partially insoluble complex medium was investigated. The goal was to make the obtained extraction samples suitable for the two-dimensional electrophoresis, thus make the further proteome analysis possible. Compared to non-denatured procedure, the denatured one, precipitating with 10% TCA in acetone, efficiently eliminated the interference substances from the cell lysate. Thiourea in the rehydration solution enhanced the resolubilization of protein pellets but led to heavy horizontal streaking in the 2-DE gels. High protein loading amount improved the resolution of some low abundance proteins but did not adapt to the high abundance proteins. And it was also important to collect cells at appropriate culture time according to the analysis target. With the optimized protein extraction protocol, the protein expression patterns of S. avermitilis during the onset of avermectin production in complex medium were analyzed.