略论农村思想政治教育实效载体的模式建构

农村思想政治教育实效载体的模式建构应着力从横向模式、纵向模式和立体模式入手。横向模式主要有共赢发展模式、互动发展模式;纵向模式主要指传统载体向现代载体、后现代载体的转化和重塑;立体模式主要指和谐维、节约维和实效维。

大数据视域下高校精准资助模式创新与载体建构

当前高校学生资助工作存在着资助工作缺乏时效性、资助过程缺乏公平性、资助方式缺乏针对性、资助育人成果缺乏成效性等问题。运用大数据驱动高校的资助工作,能够实现精准识别、精准定制、精准滴灌、精准评估,从而实现从传统资助向精准资助的转型升级。高校精准资助工作由数据采集与分析系统、科学研判与决策系统、动态评估与实时反馈系统、管理协同与制度保障系统组成。

Construction of a RNA Interference Vector of Brassica napus L.Pyruvate Kinase Gene

A predominantly expressed gene, pyruvate kinease （PK） gene, control ing oil accumulation, had been identified in previous study. To construct a PK RNAi vector, a 498-bp target PK gene sequence was amplified and transferred into the pEASY-T1vector. Subsequently, the target DNA fragments were cut off by enzymes Not I and Xho I and directional y inserted into plant RNAi platform vector pHurricane, a newly developed RNAi vector in our lab, to form the PK inverse repeats. The PK inverted repeats fragment was then cloned into a modified vector pCAMBIA1390, driven by a rapeseed seed-specific promoter napin. Restriction enzyme digestion verified the successful construction of RNA interference vector. The PK RNAi con-struction wil lay a foundation for study in the function of PK in oil accumulation and metabolism in rapeseeds.

Cloning of Broad-spectrum Anti-disease NPR1 Gene with RT-PCR and Construction of Its Protein Expression Vector

[Objective] It is to clone broad-spectrum anti-disease gene NPR1 and to construct its protein expression vector.[Method] First to extract total RNA of Arabidopsis thaliana and design relevant primers,and then the method of reverse transcription PCR was adopted to clone.With the method of enzyme digestion and ligation,this gene will be directed into protein expression vector.[Result] After relevant testing,NPR1 was inserted into vector pMXB10 to obtain pMXB10-NPR1 protein expression vector.[Conclusion] Protein expression vector including NPR1 was successfully constructed.

Construction of Plant-based Expression Vector of Polyphosphate Kinase Gene

[Objective] The aim was to construct the fusion expression vector of polyphosphate kinase（PPK） and green fluorescent protein（GFP） genes.[Method] In this study,the primers were designed based on PPK gene sequence（L03719） of E.coli DH5α in Genbank.Genomic DNA of E.coli DH 5α was extracted as template for the amplification of PPK gene by PCR method.By using In-Fusion@ HD Cloning Kit,the PPK gene was directionally cloned into NcoI site of the pCAMBIA1302 vector.[Result] Sequencing results showed that the 2.0 kb long fragment of PPK gene was inserted into the plant-based expression vector pCAMBIA1302 in front of GFP gene.[Conclusion] The fusion expression vector of PPK and GFP genes were successfully constructed.

Construction of Plant Expression Vector for Recombinant Human Epidermal Growth Factor (hEGF)

ObjectiveThis study aimed to construct plant expression vector for recombinant human epidermal growth factor （hEGF） and further to provide a basis for the expression of hEGF in peanut hairy root system. MethodAccording to the hEGF sequence in GenBank, hEGF was synthesized artificially; subsequently, hEGF gene was ligated with green fluorescent protein （GFP） gene, and their ligation product was then amplified with primers flanked with corresponding endonuclease cleavage sites, followed by double digestion by Sal I and EcoR I of the amplified products; next, pRI 101 AN DNA was extracted and digested by both Sal I and EcoR I; susequently, the digestion products of hEGF and GFP ligation fragment by Sal I and EcoR I and the digestion products of pRI 101 AN plasmid DNA by Sal I and EcoR I were ligated, and their ligation product was transformed into Escherichia coli XL10-Gold, followed by extraction of DNA from the recombinants exhibiting green fluorescence, which was then identified by enzymatic digestion and PCR, and the verified recombinant plasmid DNA was named pBZG101. ResultHuman epidermal growth factor gene （hEGF） and green fluorescent protein gene （GFP） were successfully ligated, and their ligation fragment was successfully ligated to pRI 101 AN DNA, finally with the acquirement of the plant expression vector for recombinant human epidermal growth factor-（pBZG101）. ConclusionThe plant expression vector for recombinant human epidermal growth factor-（pBZG101）- was successfully constructed in this study.

中西方环境意识比较

中西方环境意识思想存在着显著差异。通过从联系紧密的三个面向——环境意识概念、不同学科影响和环境意识建构载体进行比较,研究发现:不同预设的中西方环境意识概念差别较大,西方介入环境意识探讨和推动的学科接续性和贯通性较好,中国则出现了学科的无法接续,环境意识建构与社会进程、社会制度和公民社会成长关系密切。

Construction of Plant Virus Expression Vector pClYVV/CP/W and Expression of GFP

[ Objective] The study was to report the construction of plant virus expression vector pCIYVV/CP/W and the expression of green fluorescent protein（GFP） with pCIYVV/CP/W, and to develop effective plat virus vector for plant bioreactor to produce useful protein. [ Method] A section of multiple cloning sites among NIb/CP genes in pCIYVV genome and deoxyribonucleotide polylinker of cleavage recognition sequence containing viral protease Nla were cloned with infectivity full-length cDNA of clover yellow vein virus （CIYVV）, and pCIYVV/CP/W vector was constructed, GFP gene was inserted into pCIyVV/CP/W to construct the pCIYVV/CP/W/GFP vector. The transcription situation of recombinant virus clone was detected by RT-PCR, and targeted gene products expressed by recombinant virus clone were detected with western blot （WB）. [Result] The broad bean seedling inoculated with pCIYVV/CP/W/GFP expressed the same symptom as wild type CIYVV, morbidity was of 100%, the result showed that recombinant virus clone pCIYVV/CP/W/GFP didn＇t suppress, insertion of foreign gene didn＇t destroy the open reading frame of pCIYVV/CP/W. Foreign gene can keep living in F, progeny virus genorne steadily, recombinant virus clone pCIYVV/CP/W/GFP could steadily express GFP in progeny virus at least.[ Conclusion] The useful plant virus vector was provided for useful protein expressing.

Construction of RNAi Vector Which Resist Cucumber Mosaic Virus and Transformation of Tobacco

[Objective] Aimed to construct RNAi vector resistant to cucumber mosaic virus and transferred this vector into tobacco. [Method] RT-PCR method was used to amplify cucumber mosaic virus NS04 and process RNA2 gene sequen of tomato isolates. The analysis results of phylogenetic tree demonstrated that the sequence in RNA2 encoded CMV-2a had 98.0% and 96.5% homology with nucleotide and amino acid of DQ412731 isolate of Zhejiang,China. The replicase fragment in CMV RAN2 gene was taken as target sequence to construct pBi35SCR2 eukaryotic expression vector,then the expression vector was identified. Through agrobacterium-mediated method,the expression vector was transferred into tabacco and PCR method was used to check the transfer. The PCR results demonstrated that the experiment had successfully construct eukaryotic expression vector of pBi35SCR2 and the expression vector was successfully transferred into tabacco. [Conclusion] The obtained transgenic tobacco could be used as challenge test material in following experiment and provided foundation for studying processing tomato resist cucumber mosaic virus.

KISS-1 inhibits the proliferation and invasion of gastric carcinoma cells

AIM:To investigate the function of the KISS-1 gene in gastric carcinoma cells and to explore its potential mechanism.METHODS:A KISS-1 eukaryotic expression vector was constructed and transfected into BGC-823 cells.Resistant clones were obtained through G418 selection.reverse transcription-polymerase chain reaction and western blotting were used to detect KISS-1 and matrix metalloproteinase-9(MMP-9)expression in transfected cells.The growth of transfected cells was investigated by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide(MTT)proliferation assays,and the cells'invasive potential was analyzed by basement membrane(Matrigel)invasion assays.The anti-tumor effects of KISS-1 were tested in vivo using allografts in nude mice.RESULTS:The expression level of KISS-1 mRNA andprotein in BGC-823/KISS-1 transfected cells were significantly higher than in BGC-823/pcDNA3.1 transfected cells(P<0.05)or the parental BGC-823 cell line(P< 0.05).The expression level of MMP-9 mRNA and protein in BGC-823/KISS-1 were significantly less than in BGC-823/pcDNA3.1(P<0.05)or BGC-823 cells(P< 0.05).MTT growth assays show the proliferation of BGC-823/KISS-1 cells at 48 h(0.642±0.130)and 72 h(0.530±0.164)were significantly reduced compared to BGC-823/pcDNA3.1(0.750±0.163,0.645±0.140)(P<0.05)and BGC-823 cells(0.782±0.137,0.685± 0.111)(P<0.05).Invasion assays indicate the invasive potential of BGC-823/KISS-1 cells(16.50±14.88)is significantly reduced compared to BGC-823/pcDNA3.1(20.22±14.87)(P<0.05)and BGC-823 cells after 24 h(22.12±16.12)(P<0.05).In vivo studies demonstrate the rate of pcDNA3.1-KISS-1 tumor growth is significantly slower than pcDNA3.1 and control cell tumor growth in nude mice.Furthermore,tumor volume of pcDNA3.1-KISS-1 tumors(939.38±82.08 mm3)was significantly less than pcDNA3.1(1250.46±44.36 mm3) and control tumors(1284.36±55.26 mm3)(P<0.05).Moreover,the tumor mass of pcDNA3.1-KISS-1 tumors(0.494±0.84 g)was significantly less than pcDNA3.1(0.668±0.55 g)and control tumors(0.682±0.38 g)(P <0.05).CONCLUSION:KISS-1 may inhibit the proliferation and invasion of gastric carcinoma cells in vitro and in vivo through the downregulation of MMP-9.

Construction and Expression of Sugarcane UGPase cDNA Prokaryotic Expression Vector

UGPase （UDP-glucose pyrophosphorylase）, one of the primary enzymes concerned with carbohydrate metabolism, catalyzes the formation of UDPG. By inserting the UGPase cDNA fragment cloned from Saccharum officinarum into PQE-30, the prokaryotic expression vector of PQE-UGP was successfully constructed. Then the vector plasmid of PQE-UGP was transformed into host bacteria M 15 and the expression of target gene was induced by Isopropyl β-D-1-Thiogalactopyranoside （IPTG）. The research laid foundation for study on the prokaryotic expression of UGPase.

Genetic Engineering of Tobacco with Double Resistance to Both Virus and Insect

Expression vector pE14 with double resistance to virus and insect was constructed by inserting CW-cp gene and Bt-toxin gene one by into T-DNA of the same binary vector pE3. Tobacco was then transformed with Agrbacterium tumefaciens (At)GV311-SE carrying PE14. Nopaline assay and PCR amplification confirmed that both CW-cp gene and Bt-toxin gene had been introduced into tobacco genome by T-DNA of PE3. Test attack with virus and demonstrated, in some of the transgenic plants, double resistance to both infection by CMV and damage by Manduca sexta.

区域推进中小学国际理解教育的载体构建

当前中小学国际理解教育存在碎片化、泛化等问题,借鉴分子生物学中载体构建的概念,可从区域推进视角提出解决问题的基本策略,即通过解构国际理解教育外源基因信息,确立基本图式,通过课程"切点"、教师"转录"、联盟"催化酶"等路径,将国际理解教育重组到现有教育基因中,在日常教育教学中获得高水平的表达,实现国际理解教育的意义和价值。

Expression of bkt and bch genes from Haematococcus pluvialis in transgenic Chlamydomonas

β-carotene ketolase and β-carotene hydroxylase encoded by bkt and bch, respectively, are key enzymes required for astaxanthin biosynthesis in Haematococcu pluvialis 34-1n. Two expression vectors containing cDNA sequences of bkt and bch were constructed and co-transformed into cell-wall-deficient Chlamydomonas reinhardtii CC-849. Transgenic algae were screened on TAP agar plates containing 10 gg mL 1 Zeomycin. PCR-Southern analysis showed that bkt and bch were integrated into the genomes of C. reinhardtii. Transcripts of bkt and bch were further confirmed by RT-PCR-Southern analysis. Compared with the wild type, transgenic algae produced 29.04% and 30.27% more carotenoids and xanthophylls, respectively. Moreover, the transgenic algae could accumulate 34% more astaxanthin than wild type. These results indicate that foreign bkt and bch genes were successfully translated into β-carotene ketolase and β-carotene hydroxylase, which were responsible for catalyzing the biosynthesis of astaxanthin in transgenic algae.