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优化巴马香猪超排和利用crisprcas9技术建立pdx-1基因敲除猪的研究.docx
文档介绍:
CONTENTS
CoNTENTS
Abstract in Chinese ·I
Abstract in English 一III
1 Introduction 1
1.1 CRISPR/Cas system ·2
1.1.1 CRISPR study history .2
1.1.2 Mechanism of CRISPR/Cas system .3
1.1.3 Problems of gene editing technology in transgenic pigs .5
1.2 Superovulation 6
1.2.1 Features ofmammalian fcIllicle formation .6
1.2.2 Purpose and significance of superovulation study .7
1.2.3 Principle of superovulation ..8
1.2.4 Hormone of mammal follicles Occur 9
1.2.5 The affecting factors of superovulation 12
1.2.6 Progress of superovulation ·1 3
1.3 PDX-1 ..14
1.3.1 Structure ofPDX-1 14
1.3.2 Expression ofPDX-I 15
1.3.3 The role ofPDX-1 in the pancreatic development ..15
1.3.4 Value ofPDX-1 gene defect pigs .15
2 Materials and Methods ..1 6
2.1 Experimental Materials 16
2.1.1 Experimental animals and CRISPR/Cas9 system 16
2.1.2 Experimental drugs 1 6
2.1.mon reagents and preparation .1 7
2.1.4 Laboratory apparatus 1 9
2.2 Experimental method 20
2.2.1 Preparation of experimental animals .20
2.2.2 Superovulation .20
2.2.3 Embryo recovery 21
2.2.4 sgRNA design ofthe target gene .21
2.2.5 Cas9 mRNA and sgRNA synthesis in vitro .22
2.2.6 Intracytoplasmic microinjection ..22
2.2.7 Embryo Transfer .23
2.2.8 Analyzing pregnant transplant recipient ..24 2.2.9 Oocyte collection 24
2.2.10 Oocyte maturation ..24
III
万方数据
东北农业大学理学硕士学位论文
2.2.1 1 IvF .24
2.2.12 ic embryos obtained ..24
2.2.13 Identification ofPDX·1 gene knock-out pigs .25
2.2.14 Statistical analysis ..25
3 Results 。26
3.1 Superovulation dose and points ofovulation relationship 26
3.2 Superovulation dose and the number of cysts relationship ..28
3.3 Superovulation dose and the number of luteal relmionship .29
3.4 Under different seasons the influence of superovulation ..30
3.5 Differences between superovulation,IVF,ICSI and ic .32
3.6 Identification ofPDX-1 defective porcine model by intracytoplasmic microinjection... ...35
4 Discussion ..37
4.1 Superovulation ·37
4.1.1 Supero 内容来自淘豆网www.taodocs.com转载请标明出处.
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