从酵母中提取谷胱甘肽的初步研究

考察了不同提取方法对从酵母中提取谷胱甘肽（ＧＳＨ）的影响,热水抽提由于其提取收率高（９０％）,耗时短（１０ｍｉｎ）经济性强而明显优于其它提取方法,对７３２阳离子交换树脂纯化ＧＳＨ进行了初步研究,主要研究了树脂颗粒大小对提纯ＧＳＨ的影响,采用６０～８０目的树脂,ＧＳＨ收率为５７．６％,虽然比８０目以上的树脂低１０％但前者操作作所需时间只有后者的１／３。     

果园间作模式下杏树与苜蓿的根系分布特征及其土壤理化性质研究

该研究采用田间小区试验,设计杏树(Prunus armeniaca)下清耕(CK)和杏树间作紫花苜蓿(T)2个处理,实地采集测定各样地不同土层紫花苜蓿的根系生物量以及杏树的侧根系生物量,并测定土壤pH、电导率及其土壤有机质和速效氮含量,分析果园间作模式下紫花苜蓿对果树侧根系垂直分布特征及其土壤理化性质的影响,为果园间作苜蓿模式的推广提供理论依据。结果表明:(1)CK与T处理下的杏树侧根生物量在土壤中的垂直分布都主要集中在20~60cm土层,其生物量分别为750.8g和737.6g,分别占总侧根生物量的64.4%和64.5%;紫花苜蓿根系生物量分布呈倒金字塔型,且主要分布在0~40cm土层(166.3g),其中0~20cm土层的根系生物量最高(97.4g),占根系总生物量的35.8%。(2)与CK处理相比,T处理可有效增加果园表层土壤的有机质含量、速效态氮含量、硝态氮含量和铵态氮含量,其中,在0~20cm土层分别显著增加17.1%、40.8%、28.5%和40.8%,在20~40cm土层分别显著增加36.1%、23.1%、60.2%和23.8%,并显著降低了表层土壤电导率,但对土壤pH无显著影响。研究认为,杏园间作牧草紫花苜蓿虽然杏树与苜蓿根系会发生较小资源的竞争,但有利于改善林下土壤的理化性质和养分状况,能够有效促进果树的生长发育。     

高纯度大豆种子线粒体的分离

利用不连续Percoll梯度分离得到高纯度的大豆(Glycine max)种子线粒体。线粒体制剂中没有检测到胞液、过氧化物体和叶绿素的污染。线粒体的完整性达到97%以上, 在0~4 ℃下至少稳定2小时。     

Probing the production of amidated peptides following genetic and dietary copper manipulations

Amidated neuropeptides play essential roles throughout the nervous and endocrine systems. Mice lacking peptidylglycine α-amidating monooxygenase (PAM), the only enzyme capable of producing amidated peptides, are not viable. In the amidation reaction, the reactant (glycine-extended peptide) is converted into a reaction intermediate (hydroxyglycine-extended peptide) by the copper-dependent peptidylglycine-α-hydroxylating monooxygenase (PHM) domain of PAM. The hydroxyglycine-extended peptide is then converted into amidated product by the peptidyl-α-hydroxyglycine α-amidating lyase (PAL) domain of PAM. PHM and PAL are stitched together in vertebrates, but separated in some invertebrates such as Drosophila and Hydra. In addition to its luminal catalytic domains, PAM includes a cytosolic domain that can enter the nucleus following release from the membrane by γ-secretase. In this work, several glycine- and hydroxyglycine-extended peptides as well as amidated peptides were qualitatively and quantitatively assessed from pituitaries of wild-type mice and mice with a single copy of the Pam gene (PAM(+/-)) via liquid chromatography-mass spectrometry-based methods. We provide the first evidence for the presence of a peptidyl-α-hydroxyglycine in vivo, indicating that the reaction intermediate becomes free and is not handed directly from PHM to PAL in vertebrates. Wild-type mice fed a copper deficient diet and PAM(+/-) mice exhibit similar behavioral deficits. While glycine-extended reaction intermediates accumulated in the PAM(+/-) mice and reflected dietary copper availability, amidated products were far more prevalent under the conditions examined, suggesting that the behavioral deficits observed do not simply reflect a lack of amidated peptides.     

Cryo-EM structure of glycoprotein C from Crimean-Congo hemorrhagic fever virus

Crimean-Congo hemorrhagic fever virus (CCHFV) is a causative agent of serious hemorrhagic diseases in humans with high mortality rates. CCHFV glycoprotein Gc plays critical roles in mediating virus-host membrane fusion and has been studied extensively as an immunogen. However, the molecular mechanisms involved in membrane fusion and Gc-specific antibody-antigen interactions remain unresolved largely because structural information of this glycoprotein is missing. We designed a trimeric protein including most of the ectodomain region of Gc from the prototype CCHFV strain, IbAr10200, which enabled the cryo-electron microscopy structure to be solved at a resolution of 2.8 ?. The structure confirms that CCHFV Gc is a class II fusion protein. Unexpectedly, structural comparisons with other solved Gc trimers in the postfusion conformation revealed that CCHFV Gc adopted hybrid architectural features of the fusion loops from hantaviruses and domain III from phenuiviruses, suggesting a complex evolutionary pathway among these bunyaviruses. Antigenic sites on CCHFV Gc that protective neutralizing antibodies target were mapped onto the CCHFV Gc structure, providing valuable information that improved our understanding of potential neutralization mechanisms of various antibodies.     

Regulatory perspective of antibiotic biosynthesis in Streptomyces

<正>Streptomycetes are Gram-positive bacteria with high GC DNA content. They produce the most abundant secondary metabolites including over two-thirds of the clinically used antibiotics of natural origin (Barka et al., 2016), for example,the important broad-spectrum antimicrobials oxytetracycline(OTC) and chlortetracycline, which are the tetracycline antibiotics, produced by Streptomyces rimosus and Strepto-     

CCL5/CCR5 axis promotes the motility of human oral cancer cells

CCL5 (previously called RANTES) is in the CC‐chemokine family and plays a crucial role in the migration and metastasis of human cancer cells. On the other hand, the effect of CCL5 is mediated via CCR receptor. RT‐PCR and flow cytometry studies demonstrated CCR5 but not CCR1 and CCR3 mRNA in oral cancer cell lines, especially higher in those with high invasiveness (SCC4) as compared with lower levels in HSC3 cells and SCC9 cells. Stimulation of oral cancer cells with CCL5 directly increased the migration and metalloproteinase‐9 (MMP‐9) production. MMP‐9 small interfering RNA inhibited the CCL5‐induced MMP‐9 expression and thereby significantly inhibited the CCL5‐induced cell migration. Activations of phospholipase C (PLC), protein kinase Cδ (PKCδ), and NF‐κB pathways after CCL5 treatment was demonstrated, and CCL5‐induced expression of MMP‐9 and migration activity was inhibited by the specific inhibitor of PLC, PKCδ, and NF‐κB cascades. In addition, migration‐prone sublines demonstrate that cells with increasing migration ability had more expression of MMP‐9, CCL5, and CCR5. Taken together, these results indicate that CCL5/CCR5 axis enhanced migration of oral cancer cells through the increase of MMP‐9 production. J. Cell. Physiol. 220: 418–426, 2009. © 2009 Wiley‐Liss, Inc.     

鸡减蛋综合征病毒(EDSV)基因组右末端结构的分析

从中国发病鸡中分离的鸡减蛋综合征病毒（ＥｇｇＤｒｏｐＳｙｎｄｒｏｍＶｉｒｕｓ,ＥＤＳＶ）弱毒株（ＡＡ－２）,其基因组全长约为３３ｋｂ。用限制性内切酶ＨｉｎｄⅢ水解ＥＤＳＶ全基因组,构建了以ｐＢｌｕｅｓｃｒｉｐｔⅡ（ＫＳ＋）为载体的右末端片段的克隆（约４．２ｋｂ）,对其进行了序列测定和结构分析。该片段全长４１８３个碱基对（ｂｐ）,位于基因组右末端８７．３ｍ．ｕ．－１００ｍ．ｕ．。结果显示,该片段与哺乳动物腺病毒右末端Ｅ４区结构不同,与禽Ⅰ型腺病毒代表株ＣＥＬＯ右末端片段亦无同源性。本文为深入了解ＥＤＳＶ基因组结构特点,ＥＤＳＶ与其他腺病毒基因结构与功能的进化关系和ＥＤＳＶ载体的构建奠定了分子生物学基础     

合成生物制造2022

本文对2022年《生物工程学报》发表的与合成生物制造相关的综述和研究论文进行了评述,重点讨论了DNA测序、DNA合成、DNA编辑、基因表达调控和数学细胞模型等底层技术,酶的设计、改造和应用技术,化学品生物催化、氨基酸及其衍生物、有机酸、天然化合物、抗生素与活性肽、功能多糖、功能蛋白质等重要产品的生物制造技术,一碳化合物和生物质原料利用技术以及合成微生物组技术,以帮助读者从一个侧面了解合成生物制造相关技术和产业的发展情况。