Cre/loxP system The expression construct was created by cloning cDNAs encoding the selected genes and the promoter into a plasmid vector. After verification of the construct by nucleotide sequencing, the transcriptional unit was cleaved out with restriction enzymes, purified with biochemical method and used for microinjection. The offspring were initially examined by PCR genotyping. The in vivo expression was checked by chemical LacZ activity staining after crossing with a Cre-expression detection mouse line (Rosa26-Floxed LacZ). The entire process took from 1999 to 2003. The expression construct was created by cloning cDNAs encoding the selected genes and the promoter into a plasmid vector. After verification of the construct by nucleotide sequencing, the transcriptional unit was cleaved out with restriction enzymes, purified with biochemical method and used for microinjection. The offspring were initially examined by PCR genotyping. The in vivo expression was checked by crossing the mice with two Cre-reporter lines. B（1〜3か月） MuCreA contains a transcriptional unit consisting of a muscle-specific promoter (alpha-actin promoter) and Cre recombinase gene. This can be used to make an inducible knock-out by crossing with the other GM mice that have Floxed genes. MuCreA is a driver mouse that can remove Floxed genes only in skeletal muscle, but the genes to be removed are marked in the partner lines. B6;FVB-Tg(ACTA1-cre)AMcle/Rbrc B6;FVB-Tg(ACTA1-cre)AMcle/Rbrc <a href='https://brc.riken.jp/mus/pcr01386'>Genotyping protocol -PCR-</a> Necessary documents for ordering:<ol><li>Order form (<A HREF="https://mus.brc.riken.jp/ja/wp-content/uploads/form/form_4.docx">Japanese</A> / <A HREF="https://mus.brc.riken.jp/en/wp-content/uploads/form/form_b.docx">English</A>)</li><li>Category I MTA: MTA for distribution with RIKEN BRC (<A HREF="https://mus.brc.riken.jp/ja/wp-content/uploads/form/form_5.docx">Japanese</A> / <A HREF="https://mus.brc.riken.jp/en/wp-content/uploads/form/form_c.docx">English</A>)</li><li>Acceptance of responsibility for living modified organism (<A HREF="https://mus.brc.riken.jp/ja/wp-content/uploads/form/form_7.docx">Japanese</A> / <A HREF="https://mus.brc.riken.jp/en/wp-content/uploads/form/form_g.docx">English</A>)</li></ol> true B (1-3 months) Otago Univ. 条件を付加する。<br>研究成果の公表にあたって寄託者の指定する文献を引用する。Int. J. Biol. Sci., 20;6(6):546-555 (2010). University of Otago In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature(s) designated by the DEPOSITOR is requested. Int. J. Biol. Sci., 20;6(6):546-555 (2010). human alpha skeletal muscle actin promoter, phage P1 Cre recombinase, SV40 poly A signal Transgenic mouse, MuCre-A Transgenic mouse, MuCre-A MuCreA contains a transcriptional unit consisting of a muscle-specific promoter (alpha-actin promoter) and Cre recombinase gene. The mouse is used to make myogenic-specific knock-outs by crossing with mice that have a floxed gene. The MuCreA produces rapid and early cleavage of floxed genes from all differentiated myogenic cells, including cardiac myocytes and skeletal myotubes. 小石　恭子 Kyoko KOISHI RBRC01386