信号肽kozak序列n糖基化对黑曲霉表达木聚糖酶的影响生物工程专业论文.docxVIP

CONTENTSCONTENTS CONTENTS CONTENTS Abstract in(：hinese ．I Abstract in English ．．III 1．Introduction ．．1 1．1 Purpose and significance ”1 1．2 Literature review ．．1 1．2．1 Overview of signal peptides ．．1 1．2．2 Introduction ofprotein glycosylation 3 1．3 Main content ofthis research ．． ．．5 2．Materials and methods 。 ．．7 2．1 Materials 7 2．1．1 Strains and vectors ．7 2．1．2 Enzymes and major biochemical reagents ．7 2．1．3 Preparation of reagents used ·’7 2．1．4 Medium 9 2．1．5 Main laboratory instruments ．9 2．2 Experimental methods ．10 2．2．1 Extraction ofAspergillus niger genome ． “10 2．2．2 Gene cloning． ．． ． ． ． ．． ．．．10 2．2．3 Agrobacterium-mediated genetic transformation ofAspergillus niger ·14 2．2．4 identification ofhomologous recombination transformants ． · ·14 2．2．5 Expression ofxylanase gene in recombinant strain 15 2．2．6 Determination ofxylanase activity ofrecombinant strain 17 2．2．7 Enzymatic properties ofxylanase recombinant strain 17 3．Results analysis ．1 8 3．1 Cloning ofxylanase gene “18 3．1．1 Cloning ofxylanase xynBm2 gene ．18 3．1．2 Cloning ofxylanase xynBm3 gene ·19 3．1．3 Cloning ofxylanase xynBNGl gene ”19 3．1．4 Cloning ofxylanase xynBNG2 gene ”21 3．1．5 Cloning ofxylanase xynBNG3 gene ．．22 3．2 Establishment ofexpression vector ．． ．．．．． ．． ．．23 3．3 Agrobacterium．mediated genetic transformation ofAspergillus niger． --· ··26 3．3．1 PCR identification ofAgrobacterium transformants ． ．． ． ． 25 3．3．2 Method ofagrobacterium mediated transformation ofAspergillus niger ． ．． ． ．·28 III 万方数据 3．4 3．4 Identification ofhomologous recombination strains 28 3．4．1 Identification ofpositive transformants 28 3．4．2 Screening ofxyna gene homologous recombination transformation 30 3．5 Expression ofxylanase gene in recombinant strain ． ． · ”32 3．5．1 PCR Real-time detection ofxylanase ． 32 3．5．2 SDS．PAGE detection ofxylanase ． ”33 3．5．3 Enzyme activity detection ofxylanase “34 3．6 Enzymatic properties ofxylanase ． ．．