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Mammalian and Avian Transgenesis - New Approaches 2006 ed. [Kõva köide]

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  • Formaat: Hardback, 281 pages, kõrgus x laius: 235x155 mm, kaal: 679 g, 12 Illustrations, color; 36 Illustrations, black and white; XX, 281 p. 48 illus., 12 illus. in color., 1 Hardback
  • Sari: Principles and Practice
  • Ilmumisaeg: 05-Dec-2005
  • Kirjastus: Springer-Verlag Berlin and Heidelberg GmbH & Co. K
  • ISBN-10: 354028415X
  • ISBN-13: 9783540284154
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Mammalian and Avian Transgenesis - New Approaches 2006 ed.Suurem pilt
  • Formaat: Hardback, 281 pages, kõrgus x laius: 235x155 mm, kaal: 679 g, 12 Illustrations, color; 36 Illustrations, black and white; XX, 281 p. 48 illus., 12 illus. in color., 1 Hardback
  • Sari: Principles and Practice
  • Ilmumisaeg: 05-Dec-2005
  • Kirjastus: Springer-Verlag Berlin and Heidelberg GmbH & Co. K
  • ISBN-10: 354028415X
  • ISBN-13: 9783540284154
Teised raamatud teemal:
Püsilink: https://www.kriso.ee/db/9783540284154.html
Märksõnad:
Mammalian and Avian Transgenesis presents a collection of novel methods for the production of a wide range of transgenic animals. The manual focuses largely on mice, but also contains protocols for successful transgenesis in rats, cows, pigs and birds. The manual provides detailed, step-by-step protocols covering all aspects of the production of transgenic animals, including the use of lentiviral vectors in gene transfer, intracytoplasmic sperm injection, nuclear transfer, large insert transgenesis, conditional gene expression systems, the use of reporter genes in transgenesis and transgenesis in large animals and birds. The text is supplemented by superb color photos. While the focus is on newly established techniques, the fundamental methods of transgenesis are also covered for those new to the field. Thus this manual is perfectly suited for those wishing to adopt new technologies in transgenesis.

The generation of transgenic mammals, mostly rodents, has become one of the most important approaches in biological and biomedical research, and is of utmost importance in the generation of genetically improved livestock. Edited by respected scholars at the California Institute of Technology, this book provides step-by-step protocols of this important technique. The manual covers not only mice and other rodents, it also describes methods successfully used in cattle, sheep and pigs, making it extremely useful for anyone working on mammalian transgenesis.  
1 Generation of Transgenic Animals Using Lentiviral Vectors
1(22)
Carlos Lois
1.1 Introduction
1(2)
1.2 The Production of Lentiviral Vectors
3(9)
1.2.1 Design and Construction of the Lentiviral Transfer Vector
3(1)
1.2.2 Promoters
4(1)
1.2.3 Reporters
4(2)
1.2.4 Plasmid Preparation
6(1)
1.2.5 Preparation of 293T Cells
6(2)
1.2.6 Transfection
8(2)
1.2.7 Viral Concentration
10(2)
1.2.8 Virus Titration
12(1)
1.3 Production of Transgenic Mice and Rats
12(6)
1.3.1 Superovulation and Embryo Collection
13(1)
1.3.2 Delivery of Lentiviruses to Single-Cell Embryos
14(3)
1.3.3 Transfer of Embryos into Recipient Females
17(1)
1.4 Establishment of Stable Strains from Lentiviral Founder Animals
18(1)
1.4.1 Mating of Founder Animals
18(1)
1.5 Safety Guidelines for Pseudotyped Retroviruses
19(2)
References
21(2)
2 Intracytoplasmic Sperm Injection (ICSI) in the Mouse
23(18)
Ming-Wen Li, K.C. Kent Lloyd
2.1 General Description of Intracytoplasmic Sperm Injection (ICSI)
23(1)
2.2 Principles of ICSI
24(1)
2.3 Quality and Treatment of Sperm Used for ICSI
25(1)
2.4 ICSI Instrumentation
26(1)
2.5 Preparation of Microtools
27(1)
2.6 Culture Media
28(1)
2.7 ICSI Procedure
29(8)
2.7.1 Preparation of Sperm
29(1)
2.7.2 Preparation of Oocytes
29(1)
2.7.3 Injection Procedure
30(2)
2.7.4 Troubleshooting
32(3)
2.7.5 Embryo Transfer
35(2)
2.8 Transgenesis by ICSI
37(1)
2.9 Reasonable Cautions and Concerns Regarding ICSI
38(1)
References
38(3)
3 Generation of Embryonic Stem (ES) Cell-Derived Embryos and Mice by Tetraploid-Embryo Complementation
41(28)
Kevin Eggan, Rudolf Jaenisch
3.1 Introduction
41(7)
3.1.1 Advantages of Using Tetraploid Embryo Complementation
44(4)
3.2 ES Cell Culture
48(4)
3.2.1 Derivation, Culture and in vitro Gene Targeting of F1 ES Cells
48(2)
3.2.2 Sub-cloning of ES Cells to Identify 39X0 Derivatives of Targeted Cell Lines
50(1)
3.2.3 Y Chromosome Genotyping by Southern Hybridization
50(1)
3.2.4 Identification of 39X0 Subclones by PCR
51(1)
3.2.5 Karyotyping of ES Cell Lines during Serial Gene Targeting
52(1)
3.3 Production of Tetraploid Embryos
52(14)
3.3.1 Isolation and In Vitro Culture of Preimplantation Embryos
53(1)
3.3.2 Electrofusion of Two-cell Embryos
54(2)
3.3.3 Electrofusion by AC Alignment and DC Pulse
56(1)
3.3.4 Manual Alignment and DC Fusion of Two-cell Embryos
57(1)
3.3.5 Culture of Tetraploid Embryos to Blastocyst Stage
58(1)
3.3.6 Microscope Set-up for Microinjection
58(2)
3.3.7 Preparation of Microinjection Instruments for Piezo Microinjection
60(2)
3.3.8 Piezo-Micromanipulator Injection of Tetraploid Blastocysts with ES Cells
62(2)
3.3.9 Embryo Transfer to Recipient Females
64(2)
3.3.10 Cesarian Section and Cross Fostering of ES Cell Tetraploid Mice
66(1)
3.4 Final Words
66(1)
References
67(2)
4 Cloning The Laboratory Mouse by Nuclear Transfer
69(28)
Kevin Eggan, Rudolf Jaenisch
4.1 Introduction
69(2)
4.2 Factors Influencing Cloning Success
71(5)
4.2.1 Cell Cycle Status of the Donor Cell
72(1)
4.2.2 Genetic Influences on the Cloning Process
72(2)
4.2.3 Intrinsic Developmental Potential of the Donor Cell
74(1)
4.2.4 Cellular Identity of the Donor Cell
74(1)
4.2.5 Epigenetic Reprogramming after Nuclear Transfer
75(1)
4.3 Methods, Equipment and Techniques
76(9)
4.3.1 Embryo Culture Media and Common Stock Solutions
76(1)
4.3.2 Mouse Strains and Animal Husbandry
77(1)
4.3.3 Preparation of Cumulus Cells for Nuclear Transfer
77(1)
4.3.4 Preparation of Tail-tip Cells for Nuclear Transfer
77(1)
4.3.5 Culture and Preparation of ES donor cells for Nuclear Transfer
78(1)
4.3.6 Microscope Set-up
78(1)
4.3.7 Micromanipulation Instruments for Nuclear Transfer
78(1)
4.3.8 Isolation of Metaphase II Oocytes for Nuclear Transfer
79(1)
4.3.9 Enucleation of MII Oocytes
80(1)
4.3.10 Nuclear Transfer
80(1)
4.3.11 Oocyte Activation and Subsequent Culture of Cloned Embryos
80(3)
4.3.12 Derivation of Nuclear Transfer ES Cells
83(1)
4.3.13 Embryo Transfer of Cloned Embryos
84(1)
4.3.14 Cesarean Section and Cross Fostering of Cloned Animals
85(1)
4.4 Protocol for Direct Injection Nuclear Transfer
85(9)
4.4.1 Production of Embryo Culture Medium, Reagents and Mice
85(3)
4.4.2 Oocyte (Egg) Collection
88(2)
4.4.3 Enucleation
90(2)
4.4.4 Donor Nucleus Isolation
92(1)
4.4.5 Nuclear Transfer
93(1)
4.4.6 Oocyte Activation and Long-Term Culture
94(1)
References
94(3)
5 Large Insert Transgenesis
97(14)
Shiaoching Gong, Nat Heintz
5.1 Introduction
97(2)
5.2 Highly Efficient BAC Modification Based on the R6Kγ Origin of Replication
99(2)
5.3 An Approach to High Throughput Studies
101(7)
5.3.1 A Precisely Modified BAC Clone for Use in the Production of Transgenic Mice (Protocol 1)
101(4)
5.3.2 Preparation of BAC DNA by Double Acetate Precipitation and CsC1 Gradient Centrifugation (Protocol 2)
105(3)
5.3.3 Injection of BAC DNA
108(1)
5.4 Tools and Materials
108(1)
5.4.1 Materials
108(1)
5.4.2 Solutions
109(1)
References
109(2)
6 Regional and Temporal Control of Genetic Manipulation in the Mouse
111(20)
Mansuo L. Hayashi, Shigemi Hayashi
6.1 Introduction
111(1)
6.2 The Cre-loxP System
111(12)
6.2.1 Principles
111(1)
6.2.2 Applications
112(2)
6.2.3 Generating Cre lines
114(1)
6.2.4 Genotyping Cre Lines
115(1)
6.2.5 Screening Cre Lines
115(3)
6.2.6 Generating Floxed Lines
118(1)
6.2.7 RNA in situ Hybridization
119(3)
6.2.8 Tyramide Signal Amplification (TSA) for Fluorescence Immunostaining
122(1)
6.2.9 The Temporally Controllable Cre-loxP System
122(1)
6.3 The Tetracycline Regulatory System
123(5)
6.3.1 Principles
123(1)
6.3.2 Applications
124(1)
6.3.3 Generating tTA/rtTA Regulator Lines
125(1)
6.3.4 Genotyping tTA/rtTA Regulator Lines
126(1)
6.3.5 Screening tTA/rtTA Regulator Lines
126(1)
6.3.6 Generating tet-Responsive Lines
127(1)
6.3.7 Genotyping tet-Responsive Lines
127(1)
6.3.8 Screening tet-Responsive Lines
127(1)
6.4 New Directions for Regional and Temporal Gene Manipulation
128(1)
References
129(2)
7 High Resolution Gene Expression Analysis Using Reporter Genes
131(42)
Niels C. Adams, Nicholas W. Gale
7.1 Introduction
131(2)
7.2 Reporter Genes
133(8)
7.2.1 Selection of the Appropriate Reporter Gene
133(3)
7.2.2 Placental Alkaline Phosphatase
136(2)
7.2.3 LacZ (β-Gal)
138(3)
7.3 Reporter Constructs
141(1)
7.3.1 Position Effects
141(1)
7.4 Protocols
142(17)
7.4.1 Silicone Plates
142(1)
7.4.2 Dissection of Embryos for Whole-mount Staining for β-Gal or PLAP visualization
142(3)
7.4.3 Whole-mount Staining of Embryos for LacZ
145(2)
7.4.4 Whole-mount PLAP Staining of Embryos
147(1)
7.4.5 Dissection of Adult Tissue
148(3)
7.4.6 Preparing Embryos and Adult Tissues for Cryo-Sectioning
151(2)
7.4.7 Cryo-sectioning
153(2)
7.4.8 Whole-mount Adult LacZ Staining
155(1)
7.4.9 Enhancements to the Standard LacZ Staining Protocol
156(1)
7.4.10 Stock Solutions
157(2)
References
159(2)
Color Plates
161(12)
8 Nuclear Transfer in the Cow
173(22)
William A. Ritchie
8.1 Introduction
173(1)
8.2 Tools and Equipment
174(4)
8.2.1 Micromanipulation
174(1)
8.2.2 Microtools
175(1)
8.2.3 Manipulation Chamber
176(1)
8.2.4 Oocyte Handling
176(1)
8.2.5 Cell Fusion
177(1)
8.2.6 Embryo Culture
177(1)
8.2.7 Supplies
177(1)
8.3 Media and Solutions
178(5)
8.3.1 Aspiration Medium
178(1)
8.3.2 Dissection Medium (50 ml)
178(1)
8.3.3 Maturation Medium
178(1)
8.3.4 Stock solution B (250 mM NaHCO3)
178(1)
8.3.5 Stock solution C (33 mM pyruvate)
179(1)
8.3.6 Stock solution D (171 mM CaCl2.2H2O)
179(1)
8.3.7 Stock solution G (60 mM Glucose)
179(1)
8.3.8 Stock solution GLN (10 mM L-Glutamine)
179(1)
8.3.9 Stock solution H (250 mM Hepes)
180(1)
8.3.10 Stock solution K (kanamycin sulphate)
180(1)
8.3.11 Stock solution L (330 mM Na lactate)
180(1)
8.3.12 Stock solution M (MgCl2- 6H20)
180(1)
8.3.13 Stock solution S2
181(1)
8.3.14 Hepes synthetic oviduct fluid (hSOF) +Ca
181(1)
8.3.15 Hepes SOF (hSOF) -Ca
182(1)
8.3.16 Culture Medium (SOFaaBSA)
182(1)
8.3.17 Fusion Medium
183(1)
8.4 Procedure
183(8)
8.4.1 Oocyte Maturation
184(1)
8.4.2 Enucleation of Oocytes
185(3)
8.4.3 Cell Preparation
188(1)
8.4.4 Injection of cells
188(1)
8.4.5 Electrofusion
189(1)
8.4.6 Activation
190(1)
8.4.7 Culture
191(1)
8.5 Troubleshooting
191(1)
8.6 Discussion
192(1)
References
192(3)
9 Production of Transgenic Pigs by DNA Microinjection
195(26)
Robert M. Petters, Rebecca L. Krisher
9.1 Introduction
195(2)
9.1.1 In Vivo Produced Embryos
195(1)
9.1.2 In Vitro Produced Embryos
196(1)
9.2 Protocol for In Vitro Production of Pig Embryos
197(15)
9.2.1 Media, Solutions and Reagents
197(7)
9.2.2 Timeline
204(1)
9.2.3 Oocyte Collection and In Vitro Maturation (Day 1)
204(3)
9.2.4 In Vitro Fertilization
207(4)
9.2.5 In Vitro Culture
211(1)
9.3 Protocols for In Vivo Production of Pig Embryos
212(3)
9.3.1 Media, Solutions and Reagents
212(1)
9.3.2 Timeline
212(1)
9.3.3 Synchronization and Superovulation of Embryo Donors
212(1)
9.3.4 Embryo Recovery
213(2)
9.4 Production of Transgenic Pigs
215(4)
9.4.1 Centrifugation and Microinjection of Embryos
215(2)
9.4.2 Embryo Recipient Selection and Embryo Transfer
217(1)
9.4.3 Methods for Animal Identification
218(1)
References
219(2)
10 Production of Transgenic Birds Using Lentiviral Vectors 221(10)
Benjamin B. Scott, Carlos Lois
10.1 Introduction
221(1)
10.2 Overview of the Strategy
222(1)
10.3 Design of Lentiviral Vectors for Transgene Expression
222(3)
10.3.1 Promoters
222(2)
10.3.2 Transgenes
224(1)
10.4 Production of Transgenic Birds
225(4)
10.4.1 Egg Preparation
225(1)
10.4.2 Injection for the Production of Mosaic Founders (FO)
226(2)
10.4.3 Transgenic Offspring
228(1)
10.4.4 Husbandry
228(1)
References
229(2)
11 Ancillary Techniques 231(46)
Shirley Pease
11.1 Introduction
231(1)
11.2 Getting Started
231(6)
11.2.1 Microinjection Equipment
232(1)
11.2.2 Mouse Stocks for Embryo Production and Implantation
233(3)
11.2.3 Rat Stocks for Embryo Production and Implantation
236(1)
11.3 Embryo Production in Rats and Mice
237(10)
11.3.1 Preparation of Hormones and Enzymes
240(1)
11.3.2 Hormone Priming of Mice
240(1)
11.3.3 Hormone Priming of Rats
241(2)
11.3.4 Embryo Collection and Culture in Mice
243(3)
11.3.5 Embryo Collection and Culture in Rats
246(1)
11.4 Transfer of Mouse and Rat Embryos
247(11)
11.4.1 Synchronization and Implantation of Recipient Mice
247(2)
11.4.2 Oviduct Transfers in Mice: Unilateral, Infundibulum or Ampulla
249(5)
11.4.3 Uterine Transfers in Mice
254(2)
11.4.4 Synchronization and Embryo Transfer in Recipient Rats
256(1)
11.4.5 Oviduct Implants in Rats, by Infundibulum or Ampulla
257(1)
11.5 Murine ES Cells
258(9)
11.5.1 Commonly used ES Cell Lines
258(5)
11.5.2 Elementary Karyotyping
263(4)
11.6 The Establishment of Stable Strains from Genetically Altered Animals
267(7)
11.6.1 Breeding from Chimeric Mice
267(5)
11.6.2 Breeding from Lentiviral Founder Animals
272(2)
References
274(3)
Subject Index 277