{"id":579,"date":"2020-12-02T16:33:48","date_gmt":"2020-12-02T07:33:48","guid":{"rendered":"https:\/\/www.promega.co.jp\/resources\/faq\/?p=579"},"modified":"2020-12-16T17:28:08","modified_gmt":"2020-12-16T08:28:08","slug":"q210","status":"publish","type":"post","link":"https:\/\/www.promega.co.jp\/resources\/faq\/cloning\/sirna\/cap-analog\/ribo-m7g-cap-analog\/q210","title":{"rendered":"\u30e9\u30a4\u30bb\u30fc\u30c8\u3092\u4f7f\u3063\u305fin vitro\u7ffb\u8a33\u3092\u884c\u3046\u5834\u5408\u3001cap\u4ed8\u52a0\u3055\u308c\u305f\u8ee2\u5199\u7523\u7269\u306fcap\u4ed8\u52a0\u3055\u308c\u3066\u3044\u306a\u3044\u8ee2\u5199\u7523\u7269\u3088\u308a\u3082\u52b9\u7387\u7684\u306b\u7ffb\u8a33\u3055\u308c\u307e\u3059\u304b?"},"content":{"rendered":"\n

Rabbit Reticulocyte Lysate(RRL)\u3084Wheat Germ Extract(WGE)\u3067\u7ffb\u8a33\u53cd\u5fdc\u3092\u884c\u3046\u5834\u5408\u3001\u52b9\u7387\u7684\u306a\u7ffb\u8a33\u306e\u305f\u3081\u306b\u307b\u3068\u3093\u3069\u306e\u8ee2\u5199\u7523\u7269\u3067cap\u69cb\u9020\u3092\u5fc5\u8981\u3068\u3057\u307e\u305b\u3093(1,2,3)\u3002cap\u4ed8\u52a0\u3055\u308c\u3066\u3044\u306a\u3044RNA\u3067\u3082\u7ffb\u8a33\u53cd\u5fdc\u306b\u52a0\u3048\u308b\u91cf\u3092\u5897\u3084\u3059\u3053\u3068\u3067\u30bf\u30f3\u30d1\u30af\u8cea\u5408\u6210\u91cf\u306fcap\u4ed8\u52a0\u3055\u308c\u305fRNA\u3068\u540c\u7b49\u306b\u306a\u308a\u307e\u3059(4)\u3002<\/p>\n\n\n\n

RRL\u3067\u306f\u6700\u5927\u523a\u6fc0\u30ec\u30d9\u30eb\u3088\u308a20mM\u4e0a\u306e\u6fc3\u5ea6\u306eKCl\u306b\u304a\u3044\u3066\u3001\u30ad\u30e3\u30c3\u30d7\u4ed8\u52a0\u3055\u308c\u3066\u3044\u306a\u3044RNA\u304b\u3089\u306e\u30bf\u30f3\u30d1\u30af\u8cea\u5408\u6210\u306b\u6700\u9069\u306a\u6761\u4ef6\u3092\u793a\u3057\u307e\u3059(5)\u3002<\/p>\n\n\n\n

RRL\u3067\u306e\u7ffb\u8a33\u306fmRNA\u306ecap\u69cb\u9020\u306e\u5b58\u5728\u306b\u6bd4\u8f03\u7684\u5f71\u97ff\u3055\u308c\u307e\u305b\u3093\u3002\u3057\u304b\u3057\u3001\u4e00\u822c\u7684\u306b\u77e5\u3089\u308c\u3066\u3044\u308bRNA\u7d50\u5408\u30bf\u30f3\u30d1\u30af\u8cea(hnRNP A1, La, hnRNP 1\/PTB, p50)\u3092\u52a0\u3048\u308b\u3053\u3068\u306b\u3088\u308a\u3001RRL\u306b\u304a\u3051\u308b\u7ffb\u8a33\u306bcap\u69cb\u9020\u304c\u5fc5\u8981\u3068\u306a\u308b\u3053\u3068\u3092Svitkin\u3089\u306e\u7814\u7a76\u30b0\u30eb\u30fc\u30d7\u306f\u767a\u898b\u3057\u307e\u3057\u305f\u3002\u3053\u308c\u3089\u306e\u30bf\u30f3\u30d1\u30af\u8cea\u306fcap\u4ed8\u52a0\u3055\u308c\u3066\u3044\u306a\u3044mRNA\u306e\u7ffb\u8a33\u3092\u8457\u3057\u304f\u963b\u5bb3\u3057\u307e\u3059\u3002\u4e00\u65b9\u3001cap\u4ed8\u52a0\u3055\u308c\u305fRNA\u306e\u7ffb\u8a33\u306b\u306f\u5f71\u97ff\u3057\u307e\u305b\u3093(6)\u3002\u8457\u8005\u3089\u306f\u7d30\u80de\u8cea\u306b\u5b58\u5728\u3059\u308b\u3053\u308c\u3089RNA\u7d50\u5408\u30bf\u30f3\u30d1\u30af\u8cea\u306e\u6a5f\u80fd\u306e\u4e00\u3064\u3068\u3057\u3066\u30015\u2019\u672b\u7aef\u306ecap\u69cb\u9020\u3092\u4ecb\u3057\u305f\u6a5f\u69cb\u306b\u3088\u308a\u30ea\u30dc\u30bd\u30fc\u30e0\u306e\u7d50\u5408\u3092\u4fc3\u3057\u3001\u8aa4\u3063\u305f\u7ffb\u8a33\u958b\u59cb\u70b9\u304b\u3089\u306e\u958b\u59cb\u3092\u59a8\u3052\u3066\u3044\u308b\u306e\u3067\u306f\u306a\u3044\u304b\u3068\u8003\u3048\u3066\u3044\u307e\u3059\u3002<\/p>\n\n\n\n

RRL\u3068WG\u306e\u7ffb\u8a33\u30ad\u30c3\u30c8\u3067\u306f\u3001cap\u4ed8\u52a0\u3055\u308c\u305f\u8ee2\u5199\u7523\u7269\u306e\u7ffb\u8a33\u52b9\u7387\u304c\u4e0a\u304c\u308b\u3053\u3068\u3082\u3042\u308a\u307e\u3059(2,3,7)\u3002\u4f8b\u3048\u3070\u3001dihydrofolate reductase\u306emRNA\u306e\u7ffb\u8a33\u3092RRL\u3067\u884c\u3063\u305f\u5834\u5408\u3001cap\u4ed8\u52a0\u3055\u308c\u305fRNA\u304ccap\u4ed8\u52a0\u3055\u308c\u3066\u3044\u306a\u3044RNA\u306e4\uff5e8\u500d\u306e\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u767a\u73fe\u3059\u308b\u3053\u3068\u3092Resto\u3089\u306f\u89b3\u5bdf\u3057\u307e\u3057\u305f(8)\u3002(PN74-Q&A)<\/p>\n\n\n\n

  1. Dasso, M.C. and Jackson, J. J. (1989) Nucl. Acids Res.<\/em> 17, 3129.<\/li>
  2. Rabbit Reticulocyte Lysate Systems<\/em> Technical Manual #TM232, Promega Corporation.<\/li>
  3. Wheat Germ Extract<\/em> Technical Manual #TM230, Promega Corporation.<\/li>
  4. Gurevich, V.V. et. al. (1991) Anal. Biochem,<\/em> 195, 207.<\/li>
  5. Kozak, M. (1990) Nucl. Acids Res.<\/em> 18, 2828.<\/li>
  6. Svitkin, Y.V. et al. (1996) EMBO J.<\/em> 15, 7147.<\/li>
  7. RiboMAX\u2122 Large Scale RNA Production Systems – SP6, T3, T7<\/em> Technical Bulletin #TB166, Promega Corporation<\/li>
  8. Resto, E. et al. (1992) Nucl. Acids Res<\/em>. 20, 5979<\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

    Rabbit Reticulocyte Lysate(RRL)\u3084Wheat Germ Extract(WGE)\u3067\u7ffb\u8a33\u53cd\u5fdc\u3092\u884c\u3046\u5834\u5408\u3001\u52b9\u7387\u7684\u306a\u7ffb\u8a33\u306e\u305f\u3081\u306b\u307b\u3068\u3093\u3069\u306e\u8ee2\u5199\u7523\u7269\u3067cap\u69cb\u9020\u3092\u5fc5\u8981\u3068\u3057\u307e\u305b\u3093(1,2,3)\u3002cap … <\/p>\n","protected":false},"author":16,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[133],"tags":[],"_links":{"self":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/579"}],"collection":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/users\/16"}],"replies":[{"embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/comments?post=579"}],"version-history":[{"count":1,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/579\/revisions"}],"predecessor-version":[{"id":580,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/579\/revisions\/580"}],"wp:attachment":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/media?parent=579"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/categories?post=579"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/tags?post=579"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}