Genome-wide identification,classification, and expression profiling of serine esterases and other esterase-related proteins in the tobacco hornworm,Manduca sexta

Serine esterases (SEs) are hydrolases that catalyze the conversion of carboxylic esters into acids and alcohols. Lipases and carboxylesterases constitute two major groups of SEs. Although over a hundred of insect genomes are known, systematic identification and classification of SEs are rarely performed, likely due to large size and complex composition of the gene family in each species. Considering their key roles in lipid metabolism and other physiological processes, we have categorized 144 M. sexta SEs and SE homologs (SEHs), 114 of which contain a motif of GXSXG. Multiple sequence alignment and phylogenetic tree analysis have revealed 39 neutral lipases (NLs), 3 neutral lipase homologs (NLHs), 11 acidic lipases (ALs), 3 acidic lipase homologs (ALHs), a lipase-3, a triglyceride lipase, a monoglyceride lipase, a hormone-sensitive lipase, and a GDSL lipase. Eighty-three carboxylesterase genes encode 29 α-esterases (AEs), 12 AEHs (e.g., SEH4-1–3), 20 feruloyl esterases (FEs), 2 FEHs, 2 β-esterases (BEs), 2 integument esterases (IEs), 1 IEH, 4 juvenile hormone esterases, 2 acetylcholinesterases, gliotactin, 6 neuroligins, neurotactin, and an uncharacteristic esterase homolog. In addition to these GXSXG proteins, we have identified 26 phospholipases and 13 thioesterases. Expression profiling of these genes in specific tissues and stages has provided insights into their functions including digestion, detoxification, hormone processing, neurotransmission, reproduction, and developmental regulation. In summary, we have established a framework of information on SEs and related proteins in M. sexta to stimulate their research in the model species and comparative investigations in agricultural pests or disease vectors.     

Niche characteristics of rodents by diverse disturbance in Alashan desert, Inner Mongolia, China

Four diverse disturbance types, namely, farmland, rotationally grazed grassland, overgrazed grassland and forbidden grazing grassland, were identified in the Alashan desert region of Inner Mongolia. Rodents were sampled in April, July and October in 2002 and 2003 using the trap-day method. Their spatial and temporal niches among the four disturbance types were assessed using the Shannon-Wiener niche breadth index and the Pianka niche overlap index. The Shannon-Wiener niche breadth index was the highest for Allactaga sibirica and Meriones meridianus (0.925 and 0.908, respectively), intermediate for Cricetulus eversmanni, Phodopus roborovskii and Citellus dauricus (0.789, 0.782 and 0.711, respectively), and lowest for Cardiocranius paradoxus (0.003). The Pianka niche overlap indices of six-paired species were the highest and varied from 0.900 to 1.000, suggesting their spatial niche overlapped almost completely. Conversely, the spatial niche overlap index for Meriones unguiculatus, Cardiocranius paradoxus, and Salpingotus kozlovi was zero, thus indicating a complete absence of competition among them. In addition, the temporal niche breadth and overlap indices varied greatly in relation to disturbance types and season. The level of temporal niche overlap in spring and autumn was low while in summer it was high. For all disturbance types and for each season, the temporal niche overlap index of Meriones meridianus and Cricetulus barabansis was always highest but for Meriones unguiculatus and Salpingotus kozlovi always lowest. __________ Translated from Acta Ecologica Sinica, 2005, 25(10): 2,637–2,643 [译自: 生态学报]     

ARRB2 promotes colorectal cancer growth through triggering WTAP

Colorectal cancer (CRC) is one of the most lethal cancers worldwide. The expression of β-arrestin2 (β-Arr2, ARRB2) in CRC has been well investigated;however, its exact mechanism causing the cancer progression remains unclear. In this study, we discovered that the expression level of ARRB2 was significantly upregulated in CRC as compared to the normal tissues by employing the Cancer Genome Atlas (TCGA) data, western blot analysis, and immunohistochemistry. Furthermore, the level of ARRB2 was correlated with the patients’ overall survival by Kaplan–Meier analysis. The higher expression of ARRB2 promoted CRC cell growth, enhanced the cell motility, and blocked cell apoptosis, which is crucial for tumor growth. Lastly, the suppression of ARRB2 expression was enough to attenuate the progression of CRC induced by azoxymethane/dextran sodium sulfate. Interestingly, we also found that the knockdown of ARRB2 decreased several cancer pathways mediated by the expression of Wilms tumor 1 associated protein (WTAP), which led to the inhibition of cell proliferation and migration. Altogether, our results demonstrated that ARRB2 promoted the growth and migration of CRC cells by regulating the WTAP expression.     

酸性区3融合重链促进基于蛋白质内含子的双载体B域缺失型凝血因子Ⅷ转基因表达

最近的研究表明,蛋白质内含子(intein)介导的B区缺失型凝血因子Ⅷ (BDD-FⅧ)的轻链和重链剪接可顺式促进后者的分泌,而且剪接反应在细胞内、外均可发生．为进一步提高基于蛋白质内含子的双载体转BDD-FⅧ基因的功效,将具有促进重链分泌作用的位于Pro¹⁶⁴⁰～Ser¹⁶⁹⁰的酸性区3(acidic region-3,AR-3)引入重链,检验对蛋白质内含子剪接的BDD-FⅧ蛋白分泌和活性的影响．用融合蛋白内含子的附加ar-3重链(HCAR3IntN)基因和轻链(IntCLC)基因共转染培养的HEK293细胞,分别用ELISA和Coatest法定量分析分泌至培养上清中剪接BDD-FⅧ蛋白量和生物活性,并用免疫印迹观察了细胞内的BDD-FⅧ剪接．结果显示,共转HCAR3IntN和IntCLC基因细胞,分泌至上清的剪接BDD-FⅧ蛋白量和活性分别为(173±26) μg/L和(1.31±0.15) U/ml,明显高于未添加ar-3的蛋白质内含子融合重链(HCIntN)与轻链(IntCLC)基因共转染细胞[(102±12) μg/L和(0.79±0.09) U/ml],提示AR-3对蛋白质内含子剪接的BDD-FⅧ蛋白分泌和活性有明显改善作用．而且,分别转HCAR3IntN和IntCLC基因细胞混合培养后的上清中,亦检测到剪接的BDD-FⅧ蛋白和活性[(35±7) μg/L和(0.28±0.08) U/ml],表明蛋白质内含子可进行不依赖细胞机制的蛋白质剪接．另外,转基因细胞总蛋白呈现明显的可与FⅧ多克隆抗体进行反应的剪接BDD-FⅧ蛋白条带,直观地反映细胞内BDD-FⅧ的剪接．为动物模体内运用蛋白质反式剪接技术的双腺相关病毒载体(AAV)转BDD-FⅧ基因实验提供了依据．     

广东本地及移民学龄儿童饮食结构和肠道菌群差异分析

[目的]探究广东本地及移民学龄儿童的饮食结构及肠道菌群分布的差异.[方法]以广东深圳为采样点,随机抽样选取48名广东本地儿童和34名移民儿童,进行膳食问卷调查和晨便采集.采用Mann-Whitney U test分析本地及移民儿童饮食因子摄入频率的差异,并使用Illumina Miseq高通量测序技术对儿童的肠道菌群进...     

pKILLIN: a versatile positive-selection cloning vector based on the toxicity of Killin in Escherichia coli

The invention of DNA cloning over 40 years ago marked the advent of molecular biology. The technique has now become a routine practice in any modern biomedical laboratory. Although positive-selection of recombinants in DNA cloning seems to be superior to blue/white selection based on the disruption of the lacZ gene, it is rarely practiced due to its high background, lack of multiple cloning sites, and inability to express the genes of interest or purify the protein products. Here we report the creation of a new positive-selection cloning vector dubbed pKILLIN, which overcomes all of the above pitfalls. The essence behind its high cloning efficiency is the extreme toxicity and small size of the toxic domain of killin, a recently discovered p53 target gene. Insertion inactivation of killin within the multiple cloning site via either blunt- or sticky-end ligation not only serves as a highly efficient cloning trap, but also may allow any cloned genes to be expressed as His-tagged fusion proteins for subsequent purification. Thus, pKILLIN is a versatile positive-selection vector ideal for cloning PCR products, making DNA libraries, as well as routine cloning and bacterial expression of genes.     

广西海岛红树林资源的调查研究

广西海岛有红树林植物２０种,分隶１４科、１９属;按生态外貌和优势种的原则,可划分为８个群系,１１个群落．     

Transcriptome expression profiles reveal response mechanisms to drought and drought-stress mitigation mechanisms by exogenous glycine betaine in maize

Biotechnology Letters - Drought stress is one of the major abiotic stresses that limit growth, development and yield of maize crops. To better understand the responses of maize inbred lines with...     

Ssp DnaB intein大肠杆菌中介导FVIII重链和轻链的连接

研究利用intein的蛋白质反式剪接功能在大肠杆菌中对凝血VIII因子(FVIII)重链和轻链的连接作用,将B结构域大部分缺失型FVIII(BDD-FVIII)于满足剪接所需的保守性氨基酸Ser1657前断裂为重链和轻链,分别与split mini Ssp DnaB intein的106个氨基酸的N端(Int-N)和48个氨基酸的C端(Int-C)融合,构建到原核表达载体pBV220。诱导表达后SDS-PAGE分析可见预期大小的BDD-FVIII蛋白条带,Western blotting用FVIII特异性抗体证明其为剪接所产生的BDD-FVIII蛋白,表明intein可有效连接BDD-FVIII的重链和轻链。为进一步甲型血友病基因治疗研究应用intein以双腺相关病毒载体(AAV)携带FVIII基因,克服单个AAV载体的容量限制提供了依据。     

阿拉善地区不同生境小型兽类群落多样性研究

本文对内蒙古阿拉善地区不同生境中小型兽类群落的物种组成和多样性进行了初步研究。1998 年8～9 月和1999 年8～9 月, 2002 年10 月, 在8 种不同生境类型中共布放54 个样地, 22 606 个有效铗日, 捕获小型兽类标本866 只, 分属2 目6 科, 共20 种。多样性分析结果表明: 8 种不同生境的小型兽类群落Shannon-Wiener 多样性指数在1.186～1.589 之间; Pielou 均匀性指数在0.630 9～0.861 8 之间; Simpson 生态优势度指数在0.234 9～0.415 7 之间; Margalef 丰富度指数在0.931 8～2.020 之间。根据不同生境的小型兽类群落相似性指数聚类结果,可以将8 种生境的小型兽类群落大致划分为4 种类型: 耐旱型(群落Ⅴ和群落Ⅶ)、干旱型(群落Ⅰ、Ⅱ、Ⅲ)、农林交错型(群落Ⅵ和群落Ⅷ) 和森林型(群落Ⅳ) 。生境条件的差异对小型兽类群落物种组成和多样性有一定的限制作用, 而人为干扰有明显影响。