This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine β-casein/human lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in 50㎕ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was 20.9% (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 (8.8%) were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.
Human embryonic stem cells (hESCs) derived from the inner cell mass of blastocysts have the ability to renew themselves and to differentiate into cell types of all lineage. The present study was carried out to investigate whether the Wnt signaling pathway is related to maintaining self-renewal of hESCs. Glycogen Synthase Kinase 3 (GSK-3) inhibitor, BIO ((2''Z,3''E)-6-Bromoindirubin-3''-oxime) was treated to Miz-hES1 line for activation of Wnt signaling pathway. BIO-nontreated hESCs (control) and BID-treated hESCs were cultured for 5 days in the modified feeder-free system. During the culture of hESCs, differences were observed in the colony morphology between 2 groups. Controls were spread outwards whereas BIO-nontreated hESCs were clumped in the center and the differentiated cells were spreading outwards in the edges. The results of stem cell specific marker staining indicated that control were differentiated in large part whereas BIO-treated hESCs maintain self-renewal in the center of the colony. The results of lineage marker staining suggested that outer cells of the hESC colony were differentiated to the neuronal progenitor cells in both control and BIO-treated hESC. These results indicate that Wnt signaling is related to self-renewal in hESCs. In addition, control group showed higher composition of apoptotic cells (23.76%) than the BID-treated group (5.59%). These results indicate that BIO is effective on antapoptosis of hESCs.
The initial investment of rice cultivation by mechanical technique was high. This was only due to the cost of planting machine. We analysed the overall cost (without machine cost) and benefits of the two methods, it manifested that the mechanical method is very much beneficial over traditional rice cultivation technique. We observed that the requirement of soil, irriigation water, number of seeds etc. decreased in mechanical cultivation technique while net income increase by 13.07% with 80% government subsidy. Furthermore, the initial rooting was quicker, growth and development of pot raised seedlings was better than the traditional technique. Hence, this technique can give better return to the farmers with the help of government subsidy.