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March 15,2013
Volume 6,Issue 5
Pages -
Subject Area:Botany,Cell Biology,Enzymology,Applied Microbiology,Applied Biochemistry,Biochemistry

Title: Curriculum construction and cultivation of innovative talents of an exemplary elective course “Introduction to Life Sciences”

Authors: ZHU Daoli
PP: 498-502
Abstract: “Introduction to Life Sciences” is a comprehensive course, which can promote a blend of arts, sciences and other professional courses, implement quality education and promote innovation talent training. This course carries out the teacher-student interaction teaching of student-centered and teacher-guiding, and a variety of teacher-student interaction teaching methods. Using bidirectional interaction teaching methods and the flexible application of multimedia teaching methods, the developmental history of life sciences and social hot issues were discussed. Furthermore, the assessment on this public election course was implemented to deepen the teaching reform.
Keywords: other subjects of biology; Introduction to Life Science; curriculum construction; innovative talents; teaching reform

Title: Investigation on the sorghum endosperm cell development

Authors: WANG Zhong, GU Yunjie, WANG Huihui, ZHENG Yankun, WANG Lingling, LI Dongliang
PP: 476-497
Abstract: Sorghum KS-304 is the experimental material used in this paper, and its endosperm cell development is observed mainly by the combination of light microscopes, transmission electron microscopes (TEM) and scanning electron microscopes (SEM). The innovative results are as follows: 1) There is one or several occurrence centers of amyloplasts in cytoplasm of starchy endosperm cells (SEC). Occurrence center lies beside nuclear membrane. Occurrence center produces many plastid tubes of double layers of membranes. Starch is synthesized in plastid tube cytoplasm and forms starch granules. Then, the plastid tubes leave occurrence center due to starch granule’s enlargement and form amyloplasts. 2) SEC undergo nuclear deformation and degeneration during the development, while amyloplasts still grow during the nuclear deformation and degeneration. 3) Endosperm transfer cell (ETC) development has obvious characters of space and time. The endosperm epidermal cells closing to endosperm cavity are firstly differentiated into transfer cells, follwed by differentiating into the inner endosperm gradually. Cell wall thickening and wall ingrowths become less and less from epidermal transfer cells to the inner. The general extending direction of wall ingrowth is from endosperm cavity to inner endosperm. 4) Microtubules and a new organelle that consists of tubular small vesicles are involved in wall ingrowth formation. 5) Sorghum endosperm has only one layer of aleurone layer cells (ALC). ALC have a few amyloplasts besides aleurone granules and lipid bodies. These amyloplasts are formed from the stachy accumulation of the plastids originally around the nuclei. 6) The embryo surrounding region cells (ESRC) can still divide during the early stage of caryopsis. However, they degenerate gradually and their portion in the endosperm becomes less and less when caryopsis keeps developing and embryo keeps growing. Eventually, the embryo occupies the whole surrounding region.
Keywords: botany; sorghum; starchy endosperm cells; endosperm transfer cells; aleurone layer cells; embryo surrounding region cell; amyloplast

Title: Environmental response characteristics of leaf traits in the dominant farmland species Caragana korshinskii of Loess Plateau

Authors: GONG Chunmei, BAI Juan, ZHAO Qiulin
PP: 468-475
Abstract: The plant leaf traits are characterized by the evolutionary parameters of adaptation to the environments, including specific leaf area (SLA), N, P content and net CO2 assimilating rate and so on. In this paper, the system comparison analysis on leaf characteristics of Caragana korshinskii along the precipitation gradient was carried out, and the influence of water which is the main limiting factor as plant growth and development on leaf traits of Loess Plateau was determined. Through the combination of field investigation with laboratory analysis, we obtained the following results: SLA of C.korshinskii was the largest in Yangling, but it was relatively lower in the rest trial sites with the increasing drought stress. Furthermore, SLA and net CO2 assimilating rate was negatively correlated among several trail sites, and the net CO2 assimilating rate was the lowest in Yangling, followed by Huangling. However, it was relatively higher in Ansai, Yulin and Shenmu. In addition, plant N∶P value hadn’t significant changes along trial sites, and the correlation between N, P content and N∶P value was not significant in Yangling, but the N content and N∶P value were positively correlated significantly in Huangling and Yulin, and the P content was significantly negatively correlated with N∶P value in Ansai and Shenmu. The leaf traits of C.korshinskii presented unique regular changes along trial sites with the precipitation gradient.
Keywords: botany; Caragana korshinskii; Loess Plateau; leaf traits; environmental response

Title: Flora components and distribution of soil organic carbon of the wetlands in the middle reaches of the Heihe river

Authors: ZHAO Ruifeng, ZHANG Lihua, ZHAO Haili, JIANG Penghui
PP: 462-467
Abstract: The flora components, ecological characteristics and distribution of soil organic carbon (SOC) at different vegetation types in the wetland of the middle reaches of the Heihe river were studied using botany and pedology methods together with field investigation and sampling. The results showed that there were 84 plant species in the wetland of the middle reaches of the Heihe river, which belonged to 32 families and 70 genera. The species of compositae and gramineae were dominant, occupying 29.77% of the total species. Most of geographic elements of the flora embodied the feature of temperate zone. The perennial herbs were the main part of wetland plants. The content of SOC at was in the order of high herb wetland vegetation>lower herb wetland vegetation>woody wetland vegetation, and the difference was significant at 0-20 cm soil depth (P<0.05). The coefficient of variation of SOC was greateast at the lower herb wetland vegetation, middle at woody wetland vegetation and smallest at high herb wetland vegetation. The soil carbon density at 0-40 cm depth was 7.33, 5.44, 4.25 kg/m2 at high herb wetland vegetation, lower herb wetland vegetation and woody wetland vegetation, respectively. At the high herb and lower herb wetland vegetation, the SOC contents were more concentrated at the 0-10 cm depth, which accounted for 32% and 31% of the total at 0-40 cm depth. While the SOC content was larger at the soil depth of 10-20 cm (accounting for 33%) at the woody wetland vegetation.
Keywords: botany; wetlands in the arid region; flora components; soil organic carbon; carbon density; Heihe river

Title: Pygo2 promotes the amplification of breast cancer stem cells in transgenic mouse

Authors: LUO Qicong, LI Chao, YE Guodong, LI Boan
PP: 455-461
Abstract: Objective: To investigate the function of Pygo2 protein on regulating tumorigenesis of breast cancer in Pygo2-transgenic mice. Methods: K14-2譮lag-Pygo2-transgenic mice were established, and the mammogenesis, breast cancer stem cells population and incidence of breast cancer were examined. Results: The mammogenesis, percentage of breast cancer stem cell population and incidence of breast cancer in Pygo2-transgenic mice were all different from those in wild-type littermates. Conclusion: Pygo2 protein plays an important role in the process of tumorigenesis of breast cancer.
Keywords: oncology; Pygo2; transgenic mouse; breast cancer stem cell

Title: Relationship of enzymologic properties with structure features of trypsin from different animal groups

Authors: WANG Junren, JIANG Guoliang, WU Zhiqiang
PP: 443-454
Abstract: The reported trypsin sequences (EC 3.4.21.4) in Genebank were selected, and relationship between the structure features and enzymologic properties in insect trypsin (I-TRY), temperate and tropical zone crustacean trypsin (TTC-TRY), cold adapted fish trypsin (CAF-TRY), temperate and tropical zone fish trypsin (TTF-TRY) and higher vertebrate trypsin (HV-TRY) was comparied. It was found that, the evolutionary trend between the nummbers of amino acids, molecular diameter, diversity of molecular structure and fuction of arthropods and vertebrates trypsins were different in two groups with highest evolutionary position within invertebrates and chordates separately. The animal trypsin active centre and amino acid side chains round were strictly conservative. With differences of Arg, Glu, Asp, His, Cys, Pro, Ala, Tyr and Met around periphery and active centre, the stability, flexibility and peaction energy consumption of animal trypsin were significantly different. The Kcat of CAF-TRY, TTF-TRY and TTC-TRY was increased, while the Km of them was reduced. The catalytic efficiency was the following order: CAF-TRY>TTF-TRY＞TTC-TRY＞I-TRY and HV-TRY.
Keywords: enzymology; trypsin; structure feature; enzymologic property

Title: Isolation and identification of microbe producing biosurfactant

Authors: ZUO Yuhu, BI Sining, HOU Jumei, LIU Tong
PP: 438-442
Abstract: Ten microbes which can produce biosurfactant possibly were isolated from 270 Daqing oil-contaminated soils by methods of enrichment culture, subculture, hemolytic activity assay on blood agar plates and hydrolyzing oil activity on oil agar plates. Then one excellent isolate as potential biosurfactant producer was obtained by degreasing oil assay and the emulsification index (E24). This isolate was identified as Pseudomonas sp. based on its physiological characteristics and analysis of its 16S rRNA sequence and was named as BS1.
Keywords: applied microbiology; biosurfactant; blood agar plate; emulsification index; Pseudomonas sp.

Title: Preparation of microencapsulation of Conidiobolus obscurus mycelium using air-flow piercing device

Authors: CHEN Xiangbo, WANG Danqi, CHEN Chun
PP: 431-437
Abstract: This paper studied the preparation of microencapsulation of Conidiobolus obscurus mycelium using air-flow piercing device. Through the orthogonal test on sodium alginate concentrations, CaCl2 solution concentration, mycelium dilution and pin types and air-flow pressure, the results showed that optimum experimental conditions for using air-flow piercing device to prepare mycelium microcapsulation were 4% sodium alginate, 4% CaCl2, one fold dilution of mycelium, 0.05 mm pin diameter and 0.05 MPa air pressure. The data provided foundation for formulation of Entomophthoralean fungus and pratice for its biological control.
Keywords: microbiology; air-flow piercing device; microcapsulation; Conidiobolus obscurus; entomopathogenic fungi

Title: Study on topology of the Rv2869c from Mycobacterium tuberculosis

Authors: ZHU Wenjun, DAI Zhuqing, ZOU Man, XU Langlai, XU Dongqing
PP: 423-430
Abstract: Rv2869c, a membrane protein of Mycobacterium tuberculosis (MTB), which belongs to S2P (site-2 protease) family, encoded by Rv2869c gene, is a kind of intramembrane-cleaving proteases (I-Clips). In order to obtain the topology of Rv2869c, an S2P homologue of MTB, the topology of Rv2869c was firstly predicted by web-based topology prediction server. Then the topology of Rv2869c was investigated using the reporter gene from the fusion of the bacterial alkaline phosphatase (phoA) gene and green fluorescent protein (GFP) gene. The results showed that Rv2869c harbored four transmembrane helices, and the C-terminal located in the periplasm. This research provides certain theoretical base for prevention and treatment of tuberculosis (TB).
Keywords: microbiology; topology; reporter gene; Rv2869c; Mycobacterium tuberculosis

Title: In vitro & in vivo evaluation on the effects of 30 ku protein-chitosan conjugates on improving protein utilization of oral absorption

Authors: JIANG Xiaofeng, YU Fei, FANG Tengfei, LIAO Bifeng, YANG Bingfei, ZHANG Yaozhou
PP: 414-422
Abstract: In order to increase the target protein utilization degree of oral absorption, this study established the preparation process standards of 30 ku proteins and chitosan in silkworm pupa powder. 30 ku protein-chitosan conjugates was used to wrap the recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF), and this mixture was lavaged to the mice for increasing the absorption of rhGM-CSF. The conjugates was also used to wrap enhanced green fluorescence protein (EGFP) for increasing the stability of EGFP in artificial digestion juice. The ELISA detection showed that the amount of rhGM-CSF in serum when the mice was lavaged with rhGM-CSF wrapped with 30 ku protein-chitosan conjugates was 15 times higher than that with rhGM-CSF alone. Fluorescent microscope imaging showed that the stability of EGFP mixtured with 30 ku protein-chitosan conjugates was higher than EGFP alone in artificial digestion juice at at 37℃. Therefore, we concluded that silkworm pupa powder 30 ku protein-chitosan conjugates can improve protein utilization of oral absorption, and also protect the target proteins to avoid being digested in digestion juice.
Keywords: applied biochemistry; silkworm pupa powder; 30 ku proteins; chitosan; artificial digestion juice; stability

Title: Research on construction, expression and characteristics of the fluorescent sensor based on azurin

Authors: ZHANG Qian, YANG Yi
PP: 407-413
Abstract: As one of the copper blue protein family member, azurin is sensitive to oxidation and reduction. In order to further understanding the role of copper atomic as active center in azurin and the influence of the coordination of amino on the protein conformation, AZURIN gene was cloned for the construction of azurin protein restructuring plasmid, then we selected series sites nearby copper coordination center. The plasmid was fused into a red fluorescent protein mcherry whose excitation and emission wavelength is nearby 600 nm. The fusion protein was expressed in E.coli BL21 (DE3) and then was purified. It was found that the fusion protein had red fluorescence, and the fluorescence changed in different redox conditions, indicating redox sensitive fluorescence probe based on azurin was constructed successfully. This research provides an effective tool for the further exploration on azurin protein conformation of activity center and its sensitive environment.
Keywords: biochemistry; azurin; mcherry; redox; copper; fluorescent sensor

Title: Purification of recombinant human-like collagen by Ni(II)-based immobilized metal ion affinity chromatography

Authors: QIAO Yuanyuan, FAN Daidi, DENG Jianjun, HUI Junfeng, ZHU Chenhui, MA Xiaoxuan
PP: 402-406
Abstract: In this paper, the recombinant human-like collagen (RHLC) was purified by elution methods of iminazole and lower pH respectively with Ni2+ sepharose fast flow as the affinity medium. Various fractions of elution were collected and then protein purity was identified for by SDS-PAGE. The results showed that the purity and the recovery of the RHLC is the best by the lower pH elution with the buffer (20 mmol/L NaH2PO4, 0.5 mol/L NaCl, pH=4.0). The protein purity was more than 99%, and the recovery rate was 76%. The purified RHLC was electrophoretically pure with a relative molecular weight of 97 ku.
Keywords: protein engineering; immobilized metal ion affinity chromatography; recombinant human-like collagen; purification

Title: Construction and expression of a human diabody against bFGF stabilized by disulphide

Authors: JIANG Haowu, KANG Zhongkui, WANG Hong, LAO Xuejun, LV Weidong, TANG Yong, SONG Qifang, HUANG Jianfang, DENG Ning
PP: 396-401
Abstract: Objective: To construct and express a human disulphide stabilized diabody (ds-Diabody) against basic fibroblast growth factor (bFGF). Methods: Taking anti-bFGF human-derived single chain Fv antibody (ScFv) genes as template, using the methods of PCR and overlap PCR, the ds-Diabody was constructed by means of linking the heavy and light chain variable domains of antibody with short linking peptide, together with the mutation of the amino acids to cysteine residues in suitable position of the heavy and light chain variable domains of a human ScFv against bFGF. Results: The results of sequencing showed that the gene of ds-Diabody were constructed correctly. Western blotting demonstrated that disulfide bonds between the different peptide chains could be paired correctly to successfully form a dimer of two polypeptide chains. The ELISA results showed that the ds-Diabody has a higher specific binding activity with bFGF and more stability compared with ScFv. Conclusion: The human ds-Diabody against bFGF was constructed successfully, the affinity and stability of the antibody was improved based on ScFv, which may provide a basis for drug research of bFGF antibody.
Keywords: molecular biology; disulphide stabilized diabody; disulphide bond; mamalian cell; stability

Title: Application of real-time fluorescent quantitative PCR based on the Ct value comparison method to determine the relative gene expression

Authors: WEI Jieshu, YANG Jinfen
PP: 390-395
Abstract: Ct value comparison method is the commonly used method of fluorescent quantitative PCR determination for relative gene expression. In this paper, through the description of concrete operations, the process and characteristics of Ct value comparison method was described. Furthermore, this paper introduced the key steps and technical details of the Ct value comparison method for the determination of the relative gene expression such as primer design, cDNA templet preparason, reference gene screening, optimization of quantitative PCR system and analysis of experimental data. In addition, the application of Ct value comparison method was summarized and prospected, which provided detailed references for the application of Ct value comparison method in determining the relative gene expression.
Keywords: molecular biology; gene expression; review; fluorescent quantitative PCR; Ct value comparison method

Title: Neurotransmitters in the retina

Authors: LUO Jia, CHEN Dan, ZHOU Lihong, HUANG Jufang, WANG Hui
PP: 384-389
Abstract: The retina contains almost all the neurotransmitters in the brain, including: 1) amino acids neurotransmitters, such as glutamate (Glu), γ-aminobutyric acid (GABA), aspartate, glycine (Gly) and acetylcholine; 2) monoamine neurotransmitters, such as dopamine (DA) and 5-hydroxytryptamine (5-HT); 3) peptide neurotransmitters, such as cholecystokinin, enkephalin, glucagon, neurotensin, neuropeptide Y, auxin, substance P and vasoactive intestinal peptide; 4) some regulators, such as NO, retinoic acid and zinc ion. Among which, Gly, GABA, Gly, DA and 5-HT are regarded as the major neurotransmitters in retina, which play an inportant role on keeping the normal function of retina. This paper reviews the biosynthesis, metabolism, function and changes in pathological conditions of the major neurotransmitters in retina, which provides theoretical basis for the research, diagnosis and treatment of retina.
Keywords: neurobiology; retina; review; neurotransmitters

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