Summary of Research Project Results under JSPS FY2001
"Research for the Future Program"

1.Research Institution Kinki University
2.Research Area Life Sciences
3.Research Field Molecular Bioengineering of Food Animal Protein Resources
4.Term of Project FY 1997 〜 FY 2001
5.Project Number 97L00903
6.Title of Project Control of the Useful Genes by Genetic Engineering in Food Animals

7.Project Leader
Name Institution,Department Title of Position
Tohru Komano Kinki University, Faculty of Biology-Oriented Science and Technology Professor

8.Core Members

Names Institution,Department Title of Position
Kazuya Matsumoto Kinki University, Faculty of Biology-Oriented Science and Technology Associate Professor
Kazuhiro Saeki Kinki University, Faculty of Biology-Oriented Science and Technology Associate Professor

9.Summary of Research Results

A transgenic pig carrying the spinach-derived Δ12-desaturase (fad2) gene has been constructed by a recombinant DNA technique. cDNA of the fad2 gene was linked to the adipocyte P2 (aP2) promoter, which targets expression of linked genes specifically to adipose tissue. We observed that the Δ12-desaturase was expressed in the meat of the transgenic pigs and that the content of linoleic acid in fatty acids of the meat was increased. These results suggest that the plant-oriented fad2 gene can be functionally expressed in food animals, and that the expression can alter the composition of fatty acids of the meat of them.
In order to establish a method for the production of transgenic chicken, intra-testicular injections of foreign DNA were tested. An exogenous gene (green fluorecent protein: GFP), as a marker gene, was injected directly into the testis of matured male chicken without the laparotomy. Introduced DNA was detected by PCR analysis in embryo of next generation when the treated spermatozoa was inseminated into laying hens. Spermatozoa may act as a vector for transferring exogenous genes into the chicken.
Multidrug transporter MRP1 (multidrug resistance protein 1) expressed in the mammary glands plays a role in protecting milk against xenobiotics. We newly cloned bovine MRP1 cDNA from the mammary gland of lactating cow as one of the genes that influence the quality of daily milk. Functional analysis of bovine MRP1 revealed that the substrate specificity of bovine MRP1 differs from that of human MRP1 despite a high degree of similarity (90%) in amino acids between them.
The growth hormone (GH) gene from the pituitary gland of ostrich, the zag1 gene from mouse zygote functioning at the zygotic gene reactivation period, and the gene for S6 kinase stimulating cell growth have been isolated and characterized. A convenient system of the targeted gene disruption in chicken at cellular level (DT40 cells) has been constructed.

10.Key Words

(1)transgenic animal、(2)fatty acid desaturase、(3)transgenic pig
(4)transgenic chicken、(5)multidrug transporter、(6)GH
(7)S6 kinase、(8)zygotic activation、(9)targeting of specific gene