| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
1) Construction of transgene
Mouse PHBP cDNA coding region (1.6kbp) was ligated in pCAGGA vector which has cytomegalovirus enhancer, chiken β-actin promoter and rabbit β-globin 3'-flanking sequence in the sense direction and the transgene was named pCAPHGS. To confirme the expression of PHBP from the transgene, pCAPHGS was transfected in COS-7 and CHO cells using LIPOFECTAMINE2000 (Invitrogen) and the expression of PHBP was analyzed by western- blot analysis. The results indicated clearly the expression of PHBP from pCAPHGS.
2) Preparation and analysis of transgenic mouse
pCAPHGS was microinjected into the fertilized oocytes from C57BL/6 and the eggs were transplanted in the oviduct of female mouse. We prepared the genomic DNAs from the tails of newborne mice and analyzed them wheter they contained the transgene. As the result, we obtained 1 transgenic mouse (F0). In comparison of wild-type and F1 mice, we failed to detect any diffecences in body mass, out-looking, behavioral action and sex ratio, however, the number of newborn mice from F1 was less than from wild-type. In addition, homozygotes from F1 parents died within 1-2 days.
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