Transgenic Sheep and high quality wool production.pptx
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Transgenic sheep are genetically modified to enhance wool quality and yield through transgenesis. By introducing KAP and KIF genes, scientists improve fiber strength, elasticity, and fineness. This innovation revolutionizes wool production, benefiting the textile industry with superior, high-performance fibers.
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Transgenic Sheep and high quality wool production.pptx
- 1. TRANSGENIC SHEEP Ashwath B P ( 22B205) (ANIMAL BIOTECHNOLOGY( 19B005)) III YEAR , B.Tech Biotechnology PSG College of Technology 06/03/2025
- 2. What are they? A transgenic sheep is a genetically modified sheep that has been engineered to carry and express foreign genes (transgenes) in its genome. These transgenes are introduced to give the sheep new traits , such as improved wool quality, disease resistance, or production of valuable proteins for pharmaceutical purposes. Tracy was the first transgenically modified ewe created at Scotlands Roslin Institute. It produced the human protein alpha 1 antitrypsin, a pharmaceutical for the treatment of cystic fibrosis and emphysema.
- 3. The need? To produce therapeutic human proteins Improved Wool Production & Quality To help prevent genetic and infectious diseases in sheep Gene Therapy Models Meat Production Enhancement To produce milk with enhanced nutritional content
- 4. Cahaya Dewi - 2025 Enhanced wool production by Transgenesis
- 5. Wool follicle Structure In sheep with fine wool fibres, the follicle consists of ten cell types Two cell types that arise from the mesenchyme and eight cell types derived from the epithelium In sheep with Coarser fibres in addition to the mesenchymal and epithelial there is a additional layer called medulla. In fine wool Merino sheep the Cortex and Cuticle layer contribute to the structure of fibre.
- 6. MATURE WOOL FOLLICLE Cell division in the follicle bulb region Terminal Differentiation Synthesis of structural protein by gene expression Cross-linking and hardening of proteins at skin surface Shaping of fibre by inner root sheath Formation of fibre wool
- 7. Proteins Responsible for wool fibre Two main protein types are synthesised in cells of the cortex and cuticle Keratin Intermediate Filaments (KIF): 1. There are two types Type 1 KIF and Type 2 KIF 2. Provide filament structure Keratin-Associated Proteins (KAP): 1. The first type of KAPs are known as the glycine/tyrosine-rich KAP 2. The second type are the cysteine-rich KAPs The KIF proteins provide the filament structure in the cells, the KAP proteins provide a material that fills in the spaces between filament groups and helps to add strength to final structure.
- 8. Gene expression in wool follicle
- 9. Construction of Vector Promoter of the KIF Type 2 gene, K2.10 The K2.10 terminator sequence that assists correct processing of the messenger RNA (mRNA) labelled M (matrix attachment region) help prevent interference to operation of the K2.10 promoter Addition of gene coding region at cloning site( example : additional paracortical( KAP4) and cuticle cysteine rich( KAP5) )
- 10. Genetic manipulation by transgensis Zygote Collection : A newly fertilized sheep egg (zygote) at the single-cell stage is obtained. Microinjection of Transgene DNA : A small volume (12 picolitres) of DNA solution, containing 200600 copies of the transgene DNA, is injected into one of the pronuclei (usually the male pronucleus). Integration of Transgene DNA : After pronuclear fusion, some transgene DNA fragments integrate into the chromosomal DNA of the developing embryo. Embryo Transfer to Recipient Ewes: The injected embryos are implanted into surrogate (recipient) ewes to continue development. Gestation and Birth of Lambs: The ewes carry the developing lambs through a gestation period of approximately five months. Screening for Transgene Integration: DNA tests are conducted on newborn lambs to determine whether the transgene has successfully integrated into their genome.
- 11. DNA Testing by Southern Transfer Hybridization Black band at 3.65 kb (kilobases) seen in all sheep DNA lanes is the normal sheep gene The bands at 3.05 kb and 0.7 kb are bands from the transgene copies of the same gene (i.e. the transgenes added by microinjection).
- 12. RNA in situ hybridisation. To measure levels of the transgene RNA present, a definitive experiment which indicates both the site and level of transgene expression in the skin and follicles
- 13. Test of transgene activity in the follicles Involves analysis of the protein composition of the fibres that are formed in transgenic sheep in comparison with control sheep. Using 2-dimensional gel electrophoresis position of the normal KAP5 and KAP6 protein spots is indicated by the orange arrow
- 14. Examination of staples of wool using SEM
- 16. REFERENCES Bawden, C. S., Sivaprasad, A. V., Verma, P. J., Walker, S. K. and Rogers, G. E. (1995) Expression of bacterial cysteine biosynthesis genes in transgenic mice and sheep: toward a new in vivo amino acid biosynthesis pathway and improved wool growth. Transgenic Research vol 4, pp 87-104. Bawden, C.S., Powell, B.C., Walker, S.K. and G.E. Rogers (1998) Expression of an intermediate filament keratin transgene in sheep fibre alters structure. Transgenic Research vol 7, pp 273- 287 Bawden, S., The Australian Wool Education Trust, & University of New England. (2009). WOOL472/572 WOOL Biology and Metrology.