{"id":4673,"date":"2024-07-03T08:59:29","date_gmt":"2024-07-03T00:59:29","guid":{"rendered":"https:\/\/fanyiming.life\/?p=4673"},"modified":"2025-12-21T01:04:51","modified_gmt":"2025-12-20T17:04:51","slug":"copgfp","status":"publish","type":"post","link":"https:\/\/fanyiming.life\/?p=4673","title":{"rendered":"copGFP"},"content":{"rendered":"\n<p>\u6587\u732e\u4e2d\u770b\u5230\u6709\u4e00\u79cd&#8221;copGFP&#8221;\uff0c\u662f\u4e00\u79cd\u6bd4EGFP\u4fe1\u53f7\u66f4\u5f3a\uff08~1.3\u500d\uff09\u7684\u7eff\u8272\u8367\u5149\u86cb\u767d\uff0c\u6765\u6e90\u4e8ecopepods\uff08\u6861\u8db3\u7c7b\uff09\u7684Pontellina sp.\uff08\u7ec6\u89d2\u7c7b\u5c5e\uff09\u3002<\/p>\n\n\n\n<blockquote class=\"wp-block-quote is-layout-flow wp-block-quote-is-layout-flow\">\n<p>copGFP\u7684\u5c5e\u6027<\/p>\n\n\n\n<p>pGreenZeo\u8f7d\u4f53\u5305\u542b\u4e86\u7ecf\u8fc7\u4f18\u5316\u7684\u4eba\u7c7b\u5bc6\u7801\u5b50\u5168\u957fcopGFP\u57fa\u56e0\uff0c\u4ee5\u4fbf\u4eceCMV\u542f\u52a8\u5b50\u5728\u54fa\u4e73\u52a8\u7269\u7ec6\u80de\u4e2d\u9ad8\u6c34\u5e73\u8868\u8fbe\u8367\u5149\u86cb\u767d\u3002copGFP\u6807\u8bb0\u662f\u4e00\u79cd\u6765\u81ea\u6861\u8db3\u7c7b\uff08Pontellina sp.\uff09\u7684\u5929\u7136\u7eff\u8272\u5355\u4f53GFP\u7c7b\u4f3c\u86cb\u767d\u3002copGFP\u7684\u4e00\u4e2a\u72ec\u7279\u7279\u5f81\u662f\u5728\u86cb\u767d\u7684C\u7aef\u5b58\u5728\u4e00\u4e2a\u9644\u52a0\u7684\u53bb\u7a33\u5b9a\u5316\uff08ds\uff09\u80bd\u6bb5\uff0c\u8fd9\u4f7f\u5f97\u6210\u719f\u86cb\u767d\u7684\u534a\u8870\u671f\u5728\u4e0d\u9700\u989d\u5916\u8f6c\u5f55\u7684\u60c5\u51b5\u4e0b\u7f29\u77ed\u81f31\u5c0f\u65f6\u3002copGFP\u86cb\u767d\u662f\u4e00\u79cd\u65e0\u6bd2\u3001\u4e0d\u805a\u96c6\u7684\u86cb\u767d\uff0c\u5177\u6709\u5feb\u901f\u7684\u86cb\u767d\u6210\u719f\u3001\u9ad8\u7a33\u5b9a\u6027\uff08\u5728pH\u8303\u56f44-12\u5185\uff09\uff0c\u4e14\u4e0d\u9700\u8981\u4efb\u4f55\u989d\u5916\u7684\u8f85\u56e0\u5b50\u6216\u5e95\u7269\u3002copGFP\u86cb\u767d\u5177\u6709\u975e\u5e38\u660e\u4eae\u7684\u8367\u5149\uff0c\u5176\u4eae\u5ea6\u81f3\u5c11\u662f\u5e7f\u6cdb\u4f7f\u7528\u7684Aequorea victoria GFP\u7a81\u53d8\u4f53EGFP\u76841.3\u500d\u3002copGFP\u86cb\u767d\u53d1\u51fa\u7eff\u5149\u8367\u5149\uff0c\u5176\u7279\u6027\u5982\u4e0b\uff1a<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>\u53d1\u5c04\u6ce2\u957f\u6700\u5927\u503c &#8211; 502 nm<\/li>\n\n\n\n<li>\u6fc0\u53d1\u6ce2\u957f\u6700\u5927\u503c &#8211; 482 nm<\/li>\n\n\n\n<li>\u91cf\u5b50\u4ea7\u7387 &#8211; 0.6<\/li>\n\n\n\n<li>\u6d88\u5149\u7cfb\u6570 &#8211; 70,000 M^-1 cm^-1<\/li>\n<\/ul>\n\n\n\n<p>\u7531\u4e8e\u5176\u5353\u8d8a\u7684\u6027\u80fd\uff0ccopGFP\u662f\u4e00\u79cd\u4f18\u79c0\u7684\u8367\u5149\u6807\u8bb0\uff0c\u53ef\u4ee5\u4ee3\u66ffEGFP\u7528\u4e8e\u76d1\u6d4b\u6162\u75c5\u6bd2\u6784\u5efa\u4f53\u8fdb\u5165\u7ec6\u80de\u7684\u8fc7\u7a0b\u3002<\/p>\n<cite><a href=\"https:\/\/www.systembio.com\/wp\/wp-content\/uploads\/2020\/10\/Manual_pGreenFire_web-1.pdf\">Microsoft Word &#8211; Manual-pGreenFire-ver-082412_yk edited.docx (systembio.com)<\/a><\/cite><\/blockquote>\n\n\n\n<p>\u6211\u53ea\u80fd\u4ece\u8d28\u7c92\u4e0a\u5f97\u5230\u4e00\u4e2a800\u591abp\u7684\u5e8f\u5217\uff0c\u8fd9\u4e2acopGFP\u7684\u5e8f\u5217\u8ddfEGFP\u76f8\u4f3c\u5ea6\u975e\u5e38\u4f4e\uff0c\u53cc\u5e8f\u5217\u6bd4\u5bf9\u53ea\u670927.4%\u7684similarity\uff0c\u4e0a\u9762\u5f15\u6587\u4e2d\u63d0\u5230\u4e86\u5728C\u7aef\u52a0\u4e86\u4e00\u4e2a&#8221;destabilizing (ds) peptide&#8221;\uff0c\u6211\u60f3\u770b\u770bnatural copGFP\u662f\u5426\u5305\u542b\u8fd9\u4e00\u6bb5\u6765\u7740\uff0c\u4f46\u662fblastp\u548cblastn\u90fd\u5728Pontellina sp.\u4e2d\u627e\u4e0d\u5230\uff0c\u53ef\u80fd\u662f\u56e0\u4e3a\u8fd9\u4e2a\u5c5e\u592a\u51b7\u95e8\uff0c\u6ca1\u6709\u592a\u591a\u6570\u636e\u5e93\u4fe1\u606f\u3002<a href=\"https:\/\/www.science.org\/doi\/10.1126\/science.adk5518\">\u8fd9\u7bc7<\/a>\u6587\u7ae0\u7684Figure. 4B.\u4e2d\u5c06\u8fd9800\u591abp\u7684\u5e8f\u5217\u6807\u8bb0\u4e3aGFP-PEST\uff0cchatGFP\u5bf9&#8221;PEST&#8221;\u7684\u4ecb\u7ecd\u662f\uff1a<\/p>\n\n\n\n<blockquote class=\"wp-block-quote is-layout-flow wp-block-quote-is-layout-flow\">\n<p><strong>PEST Sequence<\/strong>:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>The PEST sequence is a peptide sequence rich in proline (P), glutamic acid (E), serine (S), and threonine (T). This sequence is known to target proteins for rapid degradation via the ubiquitin-proteasome pathway.<\/li>\n\n\n\n<li>Proteins containing PEST sequences are typically short-lived due to their rapid degradation.<\/li>\n<\/ul>\n<cite>chatGFP 4o<\/cite><\/blockquote>\n\n\n\n<p>\u5728NCBI\u4e2d\u4e0d\u6307\u5b9a\u7269\u79cd\u505ablastp\uff0c\u5f97\u5230\u7684\u6240\u6709query\u90fd\u53eacover\u5230\u524d\u534a\u6bb5\uff0c\u5e94\u8be5\u8bf4\u660eC\u7aef\u7684PEST\u5e8f\u5217\u662f\u8fd9\u4e2a\u8d28\u7c92\u7684\u6784\u5efa\u8005\u5355\u72ec\u52a0\u4e0a\u53bb\u7684\uff0c\u6240\u4ee5\u6211\u81ea\u5df1\u4ee5\u540e\u5c31\u628a\u8fd9\u6bb5\u5e8f\u5217\u6807\u8bb0\u4e3a&#8221;copGFP-PEST&#8221;\u4e86\uff0c\u63d0\u9192\u81ea\u5df1\u4e0d\u8981\u5fd8\u8bb0\u5b83\u4e0d\u662fEGFP\uff0c\u540c\u65f6\u534a\u8870\u671f\u53ea\u67091h\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image size-large is-resized\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"369\" src=\"https:\/\/fanym.oss-cn-beijing.aliyuncs.com\/wp-content\/uploads\/2024\/07\/20240703081617148-1024x369.png?x-oss-process=style\/default\" alt=\"\" class=\"wp-image-4675\" style=\"width:758px;height:auto\" srcset=\"https:\/\/fanym.oss-cn-beijing.aliyuncs.com\/wp-content\/uploads\/2024\/07\/20240703081617148-1024x369.png?x-oss-process=style\/default 1024w, https:\/\/fanym.oss-cn-beijing.aliyuncs.com\/wp-content\/uploads\/2024\/07\/20240703081617148-300x108.png?x-oss-process=style\/default 300w, https:\/\/fanym.oss-cn-beijing.aliyuncs.com\/wp-content\/uploads\/2024\/07\/20240703081617148-768x277.png?x-oss-process=style\/default 768w, https:\/\/fanym.oss-cn-beijing.aliyuncs.com\/wp-content\/uploads\/2024\/07\/20240703081617148-1536x554.png?x-oss-process=style\/default 1536w, https:\/\/fanym.oss-cn-beijing.aliyuncs.com\/wp-content\/uploads\/2024\/07\/20240703081617148-2048x738.png?x-oss-process=style\/default 2048w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<p>\u534a\u8870\u671f\u77ed\u610f\u5473\u7740\u8367\u5149\u4fe1\u53f7\u4e0d\u5bb9\u6613\u7d2f\u79ef\uff0c\u65b9\u4fbf\u505adynamic\u7684\u89c2\u5bdf\uff0c\u53e6\u4e00\u65b9\u9762\u8367\u5149\u4fe1\u53f7\u4e0d\u5bb9\u6613\u8fbe\u5230\u4e0a\u9650\uff08\u8fc7\u66dd\uff09\uff0c\u6240\u4ee5\u53ef\u80fd\u5bf9\u4e0d\u540c\u6837\u672c\/\u7ec6\u80de\u4e2d\u8367\u5149\u4fe1\u53f7\u5f3a\u5f31\u7684\u533a\u5206\u5ea6\u5927\u4e00\u70b9\u3002<\/p>\n\n\n\n<p>\u4ee5\u4e0b\u662fcopGFP-PEST\u7684\u6838\u9178\u5e8f\u5217\uff08\u5df2\u7ecf\u8fc7\u9002\u5408human\u7ec6\u80de\u8868\u8fbe\u7684\u5bc6\u7801\u5b50\u4f18\u5316\uff09\uff1a<\/p>\n\n\n\n<blockquote class=\"wp-block-quote is-layout-flow wp-block-quote-is-layout-flow\">\n<p>&gt;copGFP-PEST<\/p>\n\n\n\n<p>ggaggttctgcccccgccgccaagatcgagtgccgcatcaccggcaccctgaacggcgtggagttcgagctggtgggcggcggagagggcacccccgagcagggccgcatgaccaacaagatgaagagcaccaaaggcgccctgaccttcagcccctacctgctgagccacgtgatgggctacggcttctaccacttcggcacctaccccagcggctacgagaaccccttcctgcacgccatcaacaacggcggctacaccaacacccgcatcgagaagtacgaggacggcggcgtgctgcacgtgagcttcagctaccgctacgaggccggccgcgtgatcggcgacttcaaggtggtgggcaccggcttccccgaggacagcgtgatcttcaccgacaagatcatccgcagcaacgccaccgtggagcacctgcaccccatgggcgataacgtgctggtgggcagcttcgcccgcaccttcagcctgcgcgacggcggctactacagcttcgtggtggacagccacatgcacttcaagagcgccatccaccccagcatcctgcagaacgggggccccatgttcgccttccgccgcgtggaggagctgcacagcaacaccgagctgggcatcgtggagtaccagcacgccttcaagacccccatcgccttcgccagatctcgagatatcagccatggcttcccgccggcggtggcggcgcaggatgatggcacgctgcccatgtcttgtgcccaggagagcgggatggaccgtcaccctgcagcctgtgcttctgctaggatcaatgtgaccggt<\/p>\n<\/blockquote>\n\n\n\n<p>\u86cb\u767d\u5e8f\u5217\u4e3a\uff1a<\/p>\n\n\n\n<blockquote class=\"wp-block-quote is-layout-flow wp-block-quote-is-layout-flow\">\n<p>copGFP-PEST<\/p>\n\n\n\n<p>GGSAPAAKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRYEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGGYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFARSRDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINVTG<\/p>\n<\/blockquote>\n\n\n\n<p><\/p>\n","protected":false},"excerpt":{"rendered":"<p>\u8fd9\u7bc7\u7b14\u8bb0\u63a2\u8ba8\u4e86copGFP\uff0c\u4e00\u79cd\u6e90\u81ea\u6861\u8db3\u7c7b\u7684\u7eff\u8272\u8367\u5149\u86cb\u767d\u53d8\u4f53\u3002\u5176\u4eae\u5ea6\u6bd4EGFP\u9ad8\u7ea61.3\u500d\uff0c\u4e14\u901a\u8fc7C\u7aef\u878d\u5408\u7684PEST\u5e8f\u5217\u5b9e\u73b0\u4e86\u7ea61\u5c0f\u65f6\u7684\u77ed\u534a\u8870\u671f\uff0c\u9002\u7528\u4e8e\u52a8\u6001\u76d1\u6d4b\u3002\u4f5c\u8005\u5206\u6790\u4e86\u5176\u5e8f\u5217\u7279\u5f81\uff0c\u5e76\u4e0e\u5e38\u89c1EGFP\u8fdb\u884c\u4e86\u5bf9\u6bd4\u3002<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4],"tags":[],"class_list":["post-4673","post","type-post","status-publish","format-standard","hentry","category-note"],"_links":{"self":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/4673","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=4673"}],"version-history":[{"count":2,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/4673\/revisions"}],"predecessor-version":[{"id":13975,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/4673\/revisions\/13975"}],"wp:attachment":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=4673"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=4673"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=4673"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}