{"id":9929,"date":"2016-10-11T14:00:39","date_gmt":"2016-10-11T05:00:39","guid":{"rendered":"http:\/\/cell.brc.riken.jp\/ja\/?page_id=9929"},"modified":"2026-02-16T15:45:37","modified_gmt":"2026-02-16T06:45:37","slug":"htlv-1","status":"publish","type":"page","link":"http:\/\/cell.brc.riken.jp\/ja\/quality\/virus\/htlv-1","title":{"rendered":"HTLV-1\u691c\u67fb"},"content":{"rendered":"

\u30ec\u30c8\u30ed\u30a6\u30a4\u30eb\u30b9\u306e\u4e00\u7a2e\u3067\u3042\u308bHTLV-1\uff08Human T-cell leukemia virus type 1\uff1b\u30d2\u30c8T\u7d30\u80de\u767d\u8840\u75c5\u30a6\u30a4\u30eb\u30b91\u578b\uff09\u306f\u3001\u6210\u4ebaT\u7d30\u80de\u767d\u8840\u75c5\uff08adult T-cell leukemia, ATL\uff09\u306e\u539f\u56e0\u30a6\u30a4\u30eb\u30b9\u3068\u3057\u3066\u77e5\u3089\u308c\u3066\u3044\u307e\u3059\u3002 HTLV-1\u306eX\u907a\u4f1d\u5b50\u3092\u6a19\u7684\u3068\u3057\u305fprimer\u3092\u7528\u3044\u3066\u3001\u57f9\u990a\u7d30\u80de\u30b2\u30ce\u30e0\u306b\u30a4\u30f3\u30c6\u30b0\u30ec\u30fc\u30c8\u3055\u308c\u305fHTLV-1\u30d7\u30ed\u30a6\u30a4\u30eb\u30b9\u3092 PCR\u306b\u3088\u3063\u3066\u691c\u51fa\u3059\u308b\u65b9\u6cd5\u306b\u3088\u308a\u3001\u7d30\u80de\u6750\u6599\u304cHTLV-1\u306b\u6c5a\u67d3\u3055\u308c\u3066\u3044\u308b\u304b\u3069\u3046\u304b\u306e\u691c\u67fb\u3092\u884c\u3063\u3066\u304a\u308a\u307e\u3059\u3002
\n\u691c\u67fb\u5bfe\u8c61\u3068\u306a\u308b\u7d30\u80de\u6750\u6599\u306f\u3001\u30d2\u30c8\u8840\u6db2\u7cfb\u7531\u6765\u7d30\u80de\u682a\u304a\u3088\u3073\u30d2\u30c8\u30ea\u30f3\u30d1\u7403\u7cfb\u7d30\u80de\u682a\u3067\u3059\u3002<\/p>\n

1\uff0e\u6e96\u5099\u3059\u308b\u3082\u306e<\/strong>
\n(1)\u3000\u691c\u4f53<\/p>\n\n\n\n\n
\u62bd\u51fa\u3057\u305f\u30b2\u30ce\u30e0DNA\u6eb6\u6db2\uff1a<\/td>\n30ng\/\u00b5l\u4ee5\u4e0a<\/td>\n<\/tr>\n
\u62bd\u51fa\u65b9\u6cd5\uff1a<\/td>\n\u304a\u3088\u305d6cm dish 1\u679a\u5206\uff082\u00d7106\u500b\u7a0b\u5ea6\uff09\u306e\u7d30\u80de\u3092\u56de\u53ce\u3057\u3001PBS(-)\u3067\u6d17\u6d44\u3057\u305f\u7d30\u80de\u30da\u30ec\u30c3\u30c8\u304b\u3089DNeasy Blood & Tissue Kit ( QIAGEN Cat#69506 ) \u3092\u7528\u3044\u3066\u30b2\u30ce\u30e0DNA\u3092\u62bd\u51fa\u3059\u308b\u3002
\u6eb6\u51fa\u306b\u306f\u62bd\u51fa\u30ad\u30c3\u30c8\u306b\u6dfb\u4ed8\u3055\u308c\u3066\u3044\u308b\u30d0\u30c3\u30d5\u30a1\u30fc 200\u00b5l\u3092\u7528\u3044\u308b\u3002<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

(2)\u3000\u6a5f\u5668\u3068PCR\u30c1\u30e5\u30fc\u30d6<\/p>\n

    \n
  • 0.2ml PCR tube<\/li>\n
  • \u30c8\u30ec\u30fc \/ \u30ea\u30c6\u30fc\u30ca\u30fc \uff08PCR\u5bb9\u5668\u306b\u9069\u5408\u3059\u308b\u3082\u306e\uff09<\/li>\n
  • \u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc<\/li>\n
  • \u30a2\u30ac\u30ed\u30fc\u30b9\u30b2\u30eb\u96fb\u6c17\u6cf3\u52d5\u88c5\u7f6e<\/li>\n
  • \u30b2\u30eb\u30c8\u30ec\u30a4\u3001\u30b3\u30fc\u30e0<\/li>\n
  • \u30c8\u30e9\u30f3\u30b9\u30a4\u30eb\u30df\u30cd\u30fc\u30bf\u30fc<\/li>\n<\/ul>\n

    (3)\u3000PCR\u30d7\u30e9\u30a4\u30de\u30fc<\/p>\n

      \n
    • PCR\u30d7\u30e9\u30a4\u30de\u30fc\u3000\uff08\u53c2\u8003\u6587\u732e\u3000\uff1a\u3000S.Kwok et al., Blood, 72(4), 1117-1123, 1988\uff09<\/li>\n

      HTLV-1 X\u3000forward 5\u2019-CGGATACCCAGTCTACGTGT-3\u2019<\/p>\n

      HTLV-1 X\u3000reverse 5\u2019-GAGCCGATAACGCGTCCATCG-3\u2019<\/p>\n

    • \u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u30d7\u30e9\u30a4\u30de\u30fc<\/li>\n

      \u03b2-globin forward 5\u2019-GGTTGGCCAATCTACTCCCAGG-3\u2019<\/p>\n

      \u03b2-globin reverse 5\u2019-TGGTCTCCTTAAACCTGTCTTG-3\u2019<\/p>\n<\/ul>\n

      (4)\u3000\u8a66\u85ac<\/p>\n

        \n
      • Ex Taq Hot Start Version ( TaKaRa Cat#RR006A 250unit ) Buffer\u3001dNTP \u6dfb\u4ed8\n
      • 2% \u30a2\u30ac\u30ed\u30fc\u30b9\u30b2\u30eb<\/li>\n
      • \u30b5\u30a4\u30ba\u30de\u30fc\u30ab\u30fc\uff08100bp DNA Ladder\uff09<\/li>\n
      • 10 x Loading Buffer <\/li>\n
      • 10 mg\/ml EtBr Solution<\/li>\n
      • DDW<\/li>\n
      • \u967d\u6027\u5bfe\u7167 \uff1a \u904e\u53bb\u306e\u691c\u67fb\u3067\u967d\u6027\u3060\u3063\u305f\u30b2\u30ce\u30e0DNA<\/li>\n
      • \u9670\u6027\u5bfe\u7167 \uff1a \u904e\u53bb\u306e\u691c\u67fb\u3067\u9670\u6027\u3060\u3063\u305f\u30b2\u30ce\u30e0DNA<\/li>\n
      • 50 x TAE<\/li>\n\n\n\n\n\n\n\n\n
        Trizma base<\/td>\n242g<\/td>\n<\/tr>\n
        glatinal acetic acid<\/td>\n57.1ml<\/td>\n<\/tr>\n
        0.5m EDTA ( pH 8.0 )<\/td>\n100ml<\/td>\n<\/tr>\n
        Milli-Q\u6c34<\/td>\n842.9ml<\/td>\n<\/tr>\n
        \n
        \n<\/td>\n<\/tr>\n
        <\/td>\n\/ 1L<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
      • 1 x TAE<\/li>\n\n\n\n\n\n\n
        50 x TAE<\/td>\n20ml<\/td>\n<\/tr>\n
        Milli-Q\u6c34<\/td>\n980ml<\/td>\n<\/tr>\n
        \n
        \n<\/td>\n<\/tr>\n
        <\/td>\n\/ 1L<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
      • \uff11\u00b5g\/ml \u30a8\u30c1\u30b8\u30a6\u30e0\u30d6\u30ed\u30de\u30a4\u30c9\u6eb6\u6db2<\/li>\n\n\n\n\n
        10 mg\/ml EtBr solution<\/td>\n10\u00b5l<\/td>\n<\/tr>\n
        1 x TAE<\/td>\n100ml<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/ul>\n

        2\uff0e\u65b9\u6cd5<\/strong>
        \n\u203b \u30b2\u30ce\u30e0\u62bd\u51fa\u304c\u6e08\u3093\u3067\u3044\u306a\u3044\u691c\u4f53\u306f\u3001\u4e88\u3081\u30b2\u30ce\u30e0\u306e\u62bd\u51fa\u3092\u884c\u3046\u3002\u62bd\u51fa\u5f8c\u306e\u6fc3\u5ea6\u6e2c\u5b9a\u306730\u03bcg\/\u03bcl\u4ee5\u4e0a\u3067\u3042\u3063\u305f\u30b2\u30ce\u30e0DNA\u306fDDW\u306710\u500d\u5e0c\u91c8\u3057\u3066\u7528\u3044\u308b\u3002<\/p>\n

        (1)\u3000\u30b2\u30ce\u30e0DNA\uff08\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8DNA\uff09\u3092\u30df\u30ad\u30b5\u30fc\u3067\u5341\u5206\u306b\u64b9\u62cc\u3059\u308b\u3002
        \n(2)\u3000PCR mixture \u3092\u8abf\u6574\u3057\u3001\u305d\u308c\u305e\u308c\u306e\u691c\u4f53\u3092PCR\u30c1\u30e5\u30fc\u30d6\u306b\u5206\u6ce8\u3059\u308b\u3002
        \n\u3000\u3000\u8abf\u6574\u3059\u308b\u91cf\u306f\u691c\u4f53\u6570+2\uff08\u30dd\u30b8\u30c6\u30a3\u30d6\u30b3\u30f3\u30c8\u30ed\u30fc\u30ebx1\u3001\u30cd\u30ac\u30c6\u30a3\u30d6\u30b3\u30f3\u30c8\u30ed\u30fc\u30ebx1\uff09\u5206\u306e\u691c\u4f53\u5206\u3092\u7528\u610f\u3059\u308b\u3002<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
        PCR misture \u306e\u7d44\u6210<\/td>\n1\u691c\u4f53\u3042\u305f\u308a\u306e\u6db2\u91cf<\/td>\n<\/tr>\n
        \u30c6\u30f3\u30d7\u30ec\u30fc\u30c8DNA<\/td>\n5.0\u00b5l<\/td>\n<\/tr>\n
        PCR\u30d7\u30e9\u30a4\u30de\u30fc HTLV-I X FW (10pmol\/\u00b5l)<\/td>\n0.5\u00b5l<\/td>\n<\/tr>\n
        PCR\u30d7\u30e9\u30a4\u30de\u30fc HTLV-I X RV (10pmol\/\u00b5l)<\/td>\n0.5\u00b5l<\/td>\n<\/tr>\n
        \u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u30d7\u30e9\u30a4\u30de\u30fc \u03b2-globin FW (10pmol\/ml)<\/td>\n0.5\u00b5l<\/td>\n<\/tr>\n
        \u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u30d7\u30e9\u30a4\u30de\u30fc \u03b2-globin RV (10pmol\/ml)<\/td>\n0.5\u00b5l<\/td>\n<\/tr>\n
        10 x Ex Taq Buffer<\/td>\n2.5\u00b5l<\/td>\n<\/tr>\n
        2.5mM each dNTP mix<\/td>\n2.0\u00b5l<\/td>\n<\/tr>\n
        DDW<\/td>\n13.25\u00b5l<\/td>\n<\/tr>\n
        Ex Taq Hot Start Version<\/td>\n0.25\u00b5l<\/td>\n<\/tr>\n
        \n
        \n<\/td>\n<\/tr>\n
        \u7dcf\u91cf<\/td>\n25.0\u00b5l<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

        (3)\u3000\u5404\u691c\u4f53\u306e\u5165\u3063\u305fPCR\u30c1\u30e5\u30fc\u30d6\u3092\u30c8\u30ec\u30fc \/ \u30ea\u30c6\u30fc\u30ca\u30fc\u3068\u3068\u3082\u306b\u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc\u306b\u30bb\u30c3\u30c8\u3059\u308b\u3002
        \n(4)\u3000\u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc\u306b\u4e0b\u8a18\u306e\u8a2d\u5b9a\u3092\u5165\u529b\u3057\u3001\u53cd\u5fdc\u3055\u305b\u308b\u3002
        \n\u3000\u3000\u3000Stage 1 : 94\u2103\u30005\u5206 \uff081\u30b5\u30a4\u30af\u30eb\uff09<\/strong>
        \n\u3000\u3000\u3000Stage 2 : 94\u2103\u300010\u79d2\u3000\u2192\u300060\u2103\u30001\u79d2\u3000\u2192\u300072\u2103\u300015\u79d2 \uff0840\u30b5\u30a4\u30af\u30eb\uff09<\/strong>
        \n\u3000\u3000\u3000Stage 3 : 72\u2103\u30005\u5206 \uff081\u30b5\u30a4\u30af\u30eb\uff09<\/strong>
        \n\u3000\u3000\u3000Stage 4 : 4\u2103 \uff08\u4fdd\u5b58\uff09<\/strong>
        \n(5)\u3000PCR\u5897\u5e45\u691c\u4f53 10 \u00b5l \u306b 10 x Loading Buffer \u30921 \u00b5l \u52a0\u3048\u308b\u3002\u3000
        \n(6)\u30002%\u30a2\u30ac\u30ed\u30fc\u30b9\u30b2\u30eb\u3092\u4f5c\u88fd\u3057\u3001\u96fb\u6c17\u6cf3\u52d5\u3092\u884c\u3046\u3002
        \n(7)\u3000\u6cf3\u52d5\u5f8c\u306e\u30a2\u30ac\u30ed\u30fc\u30b9\u30b2\u30eb\u30921 \u00b5g\/ml \u30a8\u30c1\u30b8\u30a6\u30e0\u30d6\u30ed\u30de\u30a4\u30c9\u6eb6\u6db2\u3067\u67d3\u8272\u3092\u884c\u3046\u3002
        \n(8)\u3000\u30c8\u30e9\u30f3\u30b9\u30a4\u30eb\u30df\u30cd\u30fc\u30bf\u30fc\u306b\u3088\u308aDNA\u306e\u30d0\u30f3\u30c9\u3092\u78ba\u8a8d\u3059\u308b\u3002<\/p>\n

        \u6cf3\u52d5\u7d50\u679c\u4f8b
        \n<\/p>\n

        3\uff0e\u5224\u5b9a<\/strong>
        \n\u9670\u6027\u5bfe\u7167\u306b160bp\u306e\u30d0\u30f3\u30c9\u304c\u306a\u3044\u3053\u3068\u3001\u967d\u6027\u5bfe\u7167\u306b160bp\u306ePCR\u7523\u7269\u306e\u30d0\u30f3\u30c9\u304c\u3042\u308b\u3053\u3068\u3092\u78ba\u8a8d\u5f8c\u3001\u691c\u4f53\u306e\u5224\u5b9a\u3092\u884c\u3046\u3002\uff08\u03b2-globin\u306e262bp\u306e\u30d0\u30f3\u30c9\u3092\u78ba\u8a8d\u3057\u3001PCR\u963b\u5bb3\u304c\u306a\u304b\u3063\u305f\u3053\u3068\u3092\u78ba\u8a8d\uff09
        \n\u3000\u3000\u3000\u9670\u6027 : \u30d0\u30f3\u30c9\u304c\u306a\u3044
        \n\u3000\u3000\u3000\u967d\u6027 : 160bp\u306ePCR\u7523\u7269\u306e\u30d0\u30f3\u30c9\u304c\u3042\u308b<\/p>\n","protected":false},"excerpt":{"rendered":"

        \u30ec\u30c8\u30ed\u30a6\u30a4\u30eb\u30b9\u306e\u4e00\u7a2e\u3067\u3042\u308bHTLV-1\uff08Human T-cell leukemia virus type 1\uff1b\u30d2\u30c8T\u7d30\u80de\u767d\u8840\u75c5\u30a6\u30a4\u30eb\u30b91\u578b\uff09\u306f\u3001\u6210\u4ebaT\u7d30\u80de\u767d\u8840\u75c5\uff08adult T-cell leukemia, ATL\uff09\u306e\u539f\u56e0\u30a6\u30a4\u30eb\u30b9\u3068\u3057\u3066\u77e5\u3089\u308c\u3066\u3044\u307e\u3059\u3002 HTLV-1\u306eX\u907a\u4f1d\u5b50\u3092\u6a19\u7684\u3068\u3057\u305fprimer\u3092\u7528\u3044\u3066\u3001\u57f9\u990a\u7d30\u80de\u30b2\u30ce\u30e0\u306b\u30a4\u30f3\u30c6\u30b0\u30ec\u30fc\u30c8\u3055\u308c\u305fHTLV-1\u30d7\u30ed\u30a6\u30a4\u30eb\u30b9\u3092 PCR\u306b\u3088\u3063\u3066\u691c\u51fa\u3059\u308b\u65b9\u6cd5 […]<\/p>\n","protected":false},"author":14,"featured_media":0,"parent":9901,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_seopress_titles_title":"","_seopress_titles_desc":"","_seopress_robots_index":"","_seopress_robots_follow":"","_seopress_robots_imageindex":"","_seopress_robots_snippet":"","_seopress_robots_primary_cat":"","_seopress_robots_breadcrumbs":"","_seopress_robots_freeze_modified_date":"","_seopress_robots_custom_modified_date":"","_seopress_robots_canonical":"","_seopress_social_fb_title":"","_seopress_social_fb_desc":"","_seopress_social_fb_img":"","_seopress_social_fb_img_attachment_id":0,"_seopress_social_fb_img_width":0,"_seopress_social_fb_img_height":0,"_seopress_social_twitter_title":"","_seopress_social_twitter_desc":"","_seopress_social_twitter_img":"","_seopress_social_twitter_img_attachment_id":0,"_seopress_social_twitter_img_width":0,"_seopress_social_twitter_img_height":0,"_seopress_redirections_value":"","_seopress_redirections_enabled":"","_seopress_redirections_enabled_regex":"","_seopress_redirections_logged_status":"both","_seopress_redirections_param":"","_seopress_redirections_type":301,"_seopress_analysis_target_kw":"","_mc_calendar":[],"footnotes":"","_wp_rev_ctl_limit":""},"class_list":["post-9929","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9929","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/users\/14"}],"replies":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/comments?post=9929"}],"version-history":[{"count":5,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9929\/revisions"}],"predecessor-version":[{"id":15606,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9929\/revisions\/15606"}],"up":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9901"}],"wp:attachment":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/media?parent=9929"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}