flammeovirga pacifica wpaga1中琼胶酶基因的克隆表达 酶学性质分析及降解产物的初步研究word格式论文.docxVIP

3重组琼胶酶的酶学性质及其降解产物的初步研究 283.1实验材料283.1.1 重组琼胶酶283.1.2 主要试剂283.1.3 常用溶液283.1.4 主要仪器设备293.2实验方法303.2.1 重组琼胶酶的酶学性质的分析303.2.2 酶解产物的初步分析303.3结果与分析313.3.1 重组琼胶酶AgaP2049、AgaP4255、AgaP4383的酶学性质313.3.2 酶解产物的分析373.4讨论394重组琼胶酶AgaP4383 诱导表达条件的优化414.1实验材料414.2实验方法414.2.1 琼胶酶AgaP4383动力学参数的研究414.2.2 单因素试验优化重组琼胶酶AgaP4383诱导表达条件414.2.3 正交试验优化重组琼胶酶AgaP4383的诱导表达424.2.4 验证性实验424.3结果与分析424.3.1 动力学参数434.3.2 单因素试验优化重组琼胶酶AgaP4383诱导表达条件434.3.3正交试验优化重组琼胶酶AgaP4383的诱导表达454.3.4 验证性试验464.4讨论475琼胶寡糖酶法制备及其生物活性的初步研究 485.1实验材料485.1.1 原料及试剂485.1.2 主要仪器设备485.2实验方法485.2.1 琼胶寡糖的酶法制备485.2.2 琼胶寡糖体外清除DPPH自由基的能力495.2.3 细胞水平上抗氧化活性的研究505.2.4 琼胶寡糖对圣女果的保鲜作用515.3结果与分析535.3.1 琼胶寡糖的酶法制备535.3.2 琼胶寡糖体外清除DPPH自由基的能力535.3.3 在细胞水平上研究寡糖的抗氧化活性535.3.4 琼胶寡糖对圣女果的保鲜作用565.4讨论626总结与展望 646.1总结646.2展望64参考文献66硕士期间发表的论文 75致谢76ContentsContentsIVChineseabstract1Englishabstract2Prolegomenon4Introductionofdeepseamicroorganisms4IntroductionofGracilarialemaneiformis5Introductionofstrain FlammeovirgapacificaWPAGA16Agarase6Introductionof agarase6Theapplicationof agarase7Researchprogressofagaro-oligosaccharide8Purposeandsignificanceof this study10CloningandexpressionoftheagarasegenesinFlammeovirgapacificaWPAGA111Materials11Strainsandplasmids11Primers for cloningofthe agarasegenes11Reagents12Medium12Commonsolutions13Instruments and equipments15Databaseandsoftwares15Methods16AnalysisofthewholegenomeofFlammeovirgapacificaWPAGA1 and agarasegenesin thestrain16ExtractionofthegenomicDNA16Amplificationoftheagarasegenesandconstructionofrecombinantcloningplasmids16Constructionof recombinantexpressionplasmids17Inducibleexpression of agarase17SDSDetectingoftheagaraseactivity18Purificationofthe recombinant agarase19Resultsandanalysis20Analysisofthegenome ofFlammeovirga sp.WPAGA120Analysisofthe agarasegenes21Cloningandexpressionoftheagarasegenes24Purificationoftherecomninantagarases25Discussion 26Characterizationoftheagaraseandanalyzingoftheenzymaticproducts 28Materials 28Recombinantagarases28Reagents28Commonsolutions28In