CN117625430A - 一种激活放线菌沉默次生代谢产物合成基因簇表达的方法 - Google Patents
一种激活放线菌沉默次生代谢产物合成基因簇表达的方法 Download PDFInfo
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Abstract
本发明公开了一种激活放线菌沉默次生代谢产物合成基因簇(BGC)表达的方法,涉及微生物次生代谢产物技术领域,其技术要点为:该方法采用微生物群体感应信号分子刺激放线菌沉默基因簇,使放线菌沉默BGCs被激活;所使用的群体感应信号分子包括但不限于Farnesol、Tyrosol、3‑oxo‑C12‑HSL和Tryptophol。本发明的方法为一种新型激活放线菌沉默基因簇表达的方法,即为一种快速、高效激活不同放线菌中沉默BGCs的策略,本发明的方法将为激活自然界中大量存在的沉默BGCs宝库,深入发掘新型放线菌天然产物提供有效途径,从而为医药、农业等多领域的研发贡献更多更好的新型先导化合物。
Description
技术领域
本发明涉及微生物次生代谢产物技术领域,具体涉及一种激活放线菌沉默次生代谢产物合成基因簇表达的方法。
背景技术
放线菌(Actinomycetes)作为微生物的重要组成部分,具有复杂的形态分化机制和代谢调控网络,可以产生大量结构新颖的活性物质,是天然药用活性先导化合物的重要来源,被广泛应用于基础理论和应用研究。以放线菌中的主要类群链霉菌(Streptomyces)为例,大量的全基因组测序分析表明链霉菌含有丰富的次生代谢产物合成基因簇,目前发现了约6000种抗生素以及抗肿瘤、抗寄生虫和免疫抑制剂等药理活性物质。基因组测序结果显示,每个链霉菌属菌株的基因组中有20~40个次生代谢产物的生物合成基因簇(BGCs),远超过实际从链霉菌中分离得到的次生代谢产物的数量。比如在天蓝色链霉菌(Streptomyces coelicolor)A3(2)中发现的次生代谢产物只有放线紫红素(ACT)、钙依赖抗生素(Calcium-dependent antibiotic,CDA)、十一烷基灵菌红素(RED)和次甲基霉素(Mmy)等,而根据生物信息学分析,S.coelicolorA3(2)能够产生20种以上的次生代谢产物。其中,相当一部分原因是由于培养条件的限制导致大部分合成基因簇处于“沉默”或低表达状态从而无法获得相应的次生代谢产物。因此,如何有效激活放线菌中大量存在的沉默次生代谢产物合成基因簇、发掘新的次生代谢产物是当今微生物天然产物领域的研究热点和难点。
为了激活放线菌沉默基因簇的表达,获得更多新型活性次生代谢产物,寻找和开发激活沉默基因簇的新技术、新方法成为了该领域的热点前沿。目前,沉默BGCs常用的激活策略主要分为两种:一是在天然宿主菌中解除沉默BGCs的抑制因素,主要包括改变培养条件,共培养,编辑调控体系,引入强启动子等;二是避开原始菌中复杂的调控体系,直接将编码沉默BGCs的操纵子克隆到异源宿主中表达或重构沉默BGCs的控制元件后转移到异源宿主中。虽然这些常用策略已展现出了一定的优势,但也表现出了一定的局限性:传统的改善培养条件和共培养缺乏一种高通量筛选体系,工作量较大;利用基因操作激活沉默BGCs的策略不适用于培养困难或难于遗传操作的菌种;基于异源表达的激活策略虽避开了天然宿主的限制因素,但简单的异源宿主环境通常无法提供次生代谢产物生物合成的调控网络和代谢需求等。
研究发现一些化学物质可以抑制链霉菌中特定酶的活性,增加其生长压力,进而产生代谢变化。如Craney等发现一些小分子可通过抑制脂肪酸合成酶来改变链霉菌的次生代谢产物及提高抗生素产量,γ-Butyrolactones可以将S.hygroscopicus中抗真菌抗生素Validamycin的产量提升30%。群体感应(Quorum Sensing,QS)是一种微生物细胞之间的信息交流机制,近年来的研究发现QS不仅在同种微生物细胞交流中发挥作用,甚至在微生物细胞的跨种乃至跨界识别和交流过程中发挥作用,故QS信号分子有可能被作为相互拮抗的微生物细胞之间识别的信号分子,激活微生物拮抗相关基因用于细菌的竞争。基于这一假设,我们推测通过外源添加QS信号分子可被用于激活放线菌中的沉默BGCs基因的表达,从而为发掘大量存在的BGCs资源,促进新结构天然产物的发现有着极为重要的意义。
病原细菌和真菌感染逐年增加的发病率、致死率和不断增强的耐药性使其预防和治疗成为全球性的难题,人类即将面临无药可用的地步,挖掘新型抗菌药物和开发新的治疗策略已经迫在眉睫。基于群体感应信号分子作为药物靶点已成为解决病原菌感染的关键,正逐渐成为微生物群体感应领域研究的前沿热点,但关于使用群体感应信号分子激活放线菌沉默BGCs的相关方法还未见报道。模式真菌白色念珠菌(Candida albicans)和细菌铜绿假单胞菌(Pseudomonas aeruginosa)作为常见人体条件致病菌,是近年来研究的热点之一,其信号分子法尼醇(Farnesol)、酪醇(Tyrosol)和3-oxo-C12-HSL等长期以来已得到了广泛的研究,可作为用于检验群体感应信号分子激活放线菌BGCs表达的合适代表性信号分子。
为此,本发明旨在提供一种激活放线菌沉默次生代谢产物合成基因簇表达的新方法,以解决上述现有技术中的问题。
发明内容
本发明的目的是为了解决上述问题,提供一种激活放线菌沉默次生代谢产物合成基因簇表达的方法。
为了达到上述目的,本发明的技术方案如下:
本发明提供了一种激活放线菌沉默次生代谢产物合成基因簇表达的方法,所述方法为:采用代表性微生物群体感应信号分子刺激放线菌沉默基因簇,使放线菌沉默BGCs被激活(基因表达的激活通过qRT-PCR进行检测);其中,实验使用的群体感应信号分子包括但不限于Farnesol、Tyrosol、3-oxo-C12-HSL和Tryptophol。
进一步地,采用上述群体感应信号分子刺激供试的代表性放线菌天蓝色链霉菌A3(2)。
本发明还提供了上述所述的一种激活放线菌沉默次生代谢产物合成基因簇表达的方法的应用,将所述方法应用于激活放线菌中沉默BGCs,并应用于潜在的新抗生素的合成。
进一步地,应用于天蓝色链霉菌A3(2)生长状况研究。
本发明还提供上述所述的一种激活放线菌沉默次生代谢产物合成基因簇表达的方法的应用,将所述方法用于获得新型放线菌次生代谢产物及其编码的合成基因和蛋白。
进一步地,编码所述合成蛋白的核苷酸序列如SEQ ID NO:2所示。
SEQ ID NO:2:
1.SCO3232编码合成蛋白CDApeptide synthetase III,全长7254bp,具体核苷酸序列如下:
ATGGCCAAGTCCGGCTTGGAAGACATCCTGCCGCTGTCCCCGCTCCAGGAGGGGATGCTGTTC
CACAACCTCTTCGACGAGGAGGAACTGGACGCCTACAACGTCCAGGTCTTCATCGACCTGGAG
GGCGGCACGGACCCGGAGCGGCTGCGCCGGGCCGGGCAGGCGCTGCTGGAGCGGCACGCCA
ACCTGCGGGCCGCCTTCCGGCACGAGGGGCTCAAGCGCCCGGTGCAGCTCATCCCCCGCCGG
GTCGTCCTGCCCTGGGGCGAGGAGGACCTGTCGGGCGTCGCCGAACCGGAGCGGGAGGCGGC
GGCGGAGCGGGTCGCCGAGCGGGACCGCTGGACGCGCTTCGACCTGTCCCGGCCGCCGCTGA
TCCGCTTCACCCTGGTCCGGCTCGGCCCGGCCCGTCACCGGCTGCTGCTGACGCTCCACCACA
TCCTGGCGGACGGCTGGTCCATGCCGATCCTGCTGCGCGAGCTGATGACCCTGTACACCGTCC
ACGGCGACGGTACGGCCCTGCCCCGGGTGCGCCCCTACCGCGACTACCTGGGCTGGCTCGGCG
GCCGGGACCGGGACGCCGCCCGGCAGGCCTGGACGGAGGCGTTCGCCGGGCTCGACGCGCC
GAGCATCGTCGCCCCCGGCCGGGGCGCGCTCACCGCGGCGCCCGAGCGGATCGACTTCTCCG
AGGACGAGGCGGCGAGCGCCGCGCTGACCCGGTTCGCCCGGTCCAACGGGCTGACGGTGAAC
ACGGTGATCCAGGGCTGCTGGGGACTGGTCCTGTCCCATCTGACGGGCCGGGACGACGTGGT
GTTCGGCGTGACCGTCAGCGGCCGCCCGCCGGAGCTGCCCGGTATCGACACCATGGTCGGCCT
GTTCATGAACACCCTGCCGCTGCGGGTGCGGCTGCGGCCCGCCGAGACGCTGACCGGGTTCCT
GCGCCGGCTGCAGGGCGAGCAGGCGCGGCTCATCGACCACCAGTGGGTGGGCCTCGCGGAGA
TCCAGCGCTGGGCGGGTTCCGGCGAACTCTTCGACACGGCCATGGTGTTCGAGAACTATCCGC
TCAACTCCAGCCGCGGCCGCCCCCCGGGGGCCGCGCCGGACGCGGACCTCCCGACCGTCCTC
GGGGTCCGTTCGAAGGACCAGATGCACTACCCGCTGGGTCTGCTGGCCCTGCCCCGCGAGACC
CTGCGGTTCTCGCTGGGCTATCTGCCGCAGGTCTTCGACCCGGCGCGGGTCGAGGCGGTGATC
GCCGCGTTCCGCCGGGCGCTGCGCACCGTCCTGGACGCCCCGGACACCCGGGTCGGCGCGGT
CGCGCTGCTCGACCCGGAGGTGCGGGGCACCGTCCTGGAGAAGTGGAGCGGCTCGGACGACG
TGCGGCCCGCAGAGCGGTTCACCGACCTGTTCGAGGAGCAGGTGGCGCGCACCCCCGGCAAG
ACGGCGCTGATCGCCCCCGACGGGCGGCTGACCTACGCCGAACTCGACGCCGCCGCCAACCG
GCTGGCCCGTCGCCTGGTCGAGCTGGGCGTCGGCCCGGAACGGCACGTGGCGGTCGCGGTGG
GGCGCCGCACGGAACTGGTGGTCGGCATGCTGGCGGTGCTCAAGGCGGGCGGCGCCTATGTG
CCGGTCGACCCGGAGTACCCGCCGGACCGCATCCGCCACATGATCCAGGACGCGGACCCCGC
GCTCGTCCTCACCACCTCCGACGTGGACGACCGGATCGGCGAGGAGTGCTGCGGGCCGCTGA
CGTTCGTCATGGACGACCCCAACACCGGAACGTCGCTGGGCAGGCACTCCGGGACGGCTCTC
ACCGACGCCGACCGCGCGGCGCCGCTGCTGCCGGGCCATCCGGCGTACGTCATCTACACCTCG
GGCACGACGGGCCGCCCCAAGGGCGTCGTCGTCGAGCACCGGGCCCTGTCGGCGTTCGTGCG
GCACTGCCGGTCCTCCCAGGCGCCGGACATCTCGGGGCTGTCCGTGATGCAGGCGTCCGCGTC
CTTCGACCAGAGCGTCGGCTCGCTGCACGCCCCGCTGATCAGCGGCGGCTGCGTCCGGCTGAC
GGATCTGCGCGCCCTCGCCGAGACGGCCGGGTCCGAACCCGGCTTCCACCGTGCCACCTTCAT
GAAGGGCACCCCGTCGCACCTGGCGCTGCTGGCCACCATGCCGCCGGAGGTCGCGCCCTCGG
GCACCCTGACCCTGGGCGGCGAGGAGCTGCGCGGGGAGATCCTGGCACCCTGGCGCGAGGCC
GCGGGGGACGTGACGGTGGTGAACGTCTACGGGCCCACCGAGGCCACCGGTCACTGCCTGGA
GCACTGGATCGCCCCGGACCGCACGGTGGAGCCGGGCCCGGTGCCGATCGGCACCCCGCACG
AGGGGGTGCGGGTGTACGTCCTCGACTCCGCGCTGCGCCCGGTCGCACCGGGTCTGGACGGC
GAGGTGTACCTGGCGGGCGTGCAGCTGGCGCGCGGCTACCTGGGCCGCGGCGGGCTGACCGC
CGAGCGGTTCACGGCCGACCCGTTCGGCGCGCCGGGCAGCCGCATGTACCGCACGGGCGACG
TCGCGCACTGGAACGAAGCGGGGGAGCTGGTCTTCGCCGGGCGCGCGGACCGGCAGGTCAA
GCTGCGCGGGTACCGCATCGAGCTGGGCGAGATCGAGGCCGCCGTCGCGGGCGGGCCGGGTG
TGCGCCAGGCCGCGGTCGTCCTGCGCGAGGACCGGCCGGGCGACCAGCGGCTGGTGGCCTAC
GTCGTCCCGGACCCCGGCCACTGGGACGAGGCCGCCGCCCGTGCCCGGCTGGCCCTGTCCCT
GCCGGACTTCATGATGCCCTCGGCCTTCGTCGCCCTGGACGCCCTGCCGCTCAGCCCGAACGG
CAAGCTGGACCGGGCGGCGCTGCCCGCCCCGACGTACACCGGCCGGACCGCGGGCCGGGCAC
CGCGCACCCCCGCCGAGGAGATCCTCTGCGACCTGTACGCGGAGGTCCTGTCGCTGCCGGGCG
TCACCGTCGACGACGACTTCTTCGACCTCGGCGGTCACTCCCTGCTCGCCACCCGGCTCGTCA
GCCGGGTCCGCACCACGCTGGGGGCCGAGCTGTCCATCCGGCAGTTCTTCGAGGCGCCGACC
CCGGCCGCGCTCGCCGTGGTGCTCGCCGGGGCGGGCCGGGCGCGGGCCGCGCTCACCGCACG
GCCCCGTCCCGAGCGCCTCCCGCTCTCCTACGCCCAGCAGCGCCTGTGGTTCCTCCACCTGCT
CGAAGGGCCGAGCCCCACCTACAACATCCCGACGGTGCTCCGGCTGAGCGGACCGCTGCGCC
CGGACGCGCTGCGCGCGGCGCTGCTGGACGTGGTCGGGCGGCACGAGAGCCTGCGGACGAC
GTTCACCGAGGACGAGCGGGGGGCCCGGCAGGTGGTGCACCCCGCCGACGGGGTCCGGCCG
GTCTTCGAGACGGCCGAGTCCACGGAGGCCGACTACGAGGCCGACCTCGCACGGGCGGCCCG
GCACGCCTTCGATCTGGGCGCGGAGATCCCGGTACGGGCGCGGCTGCTGCGGCTGTCGGAGCG
GGAGCACGTCCTGCTGCTGCTCGTGCACCACATCGCCAGCGACGCCTGGTCCCGCGGCCCACT
GGCCCAGGACCTCACCGCCGCCTACACCGCCCGCTGCGCCGGCGACGCACCCGCCTGGCAGC
CCCTCCCCGTCCAGTACGCCGACTACGCCCTGTGGCAGCAGGAGATCCTCGGCGACGACACCG
ACCCCGACACCCTCGCCGGCCGCCAGCTCGCCTACTGGAAACAGCAGCTCGCCGGCCTGCCC
GAACGGCTCGACCTCCCCACCGACCGGCCCCGGCCCGCCACCGCCGACCACACCGGCGACCG
CGTCGAGTTCGCCCTCCCCGCCGACCTGCACACCCGTCTGACCGAACTCGCCCGGGCCACCGA
CACGACCCTCTACATGGTGCTCCAGGCCGCCCTCGCGACGCTGCTCACCCGGCACGGCGCCGG
TGAGGACATCCCGATCGGCACCCCCGTCGCCGGACGCACCGACGACGCCACCGACCACCTCG
TCGGCTTCTTCGTCAACACCCTCGTCCTGCGCACCGACACCAGCGGCAACCCCACCTTCCGCG
ACCTCCTCACCCGAGTCCGCGACACCGACCTCACCGCCTACACCCACCAGGACCTCCCCTTCG
AACGCCTCGTAGAAGCCCTCAACCCCACCCGCTCCCTCACCCACCACCCCCTCTTCCAGGTGG
TGCTGTCGCTGCGCAGCACCGCACCGCGGCGGGCGGACGGGGAGGGCGCGCCGGCGCTGCCG
GGCGGGCTGAGGGTGAGCGGCACGGGCGGTGCGGCGGCGACGGCGGCCAAGGTCGACCTCG
GGTTCTCGGTGACGGAGCGCCGGGCGGCCGACCACACCCCCGACGGGGTGGCGGGCGTCCTC
GACTTCCGCACCGACCTCTTCGACCGCGGCACCGCGCAGGGTCTGGTGGACCGGCTGGTGCG
GGTGCTCGCCGACGCGGCCGCCCACCCCGACCGGCCGCTGAGCCGCATCGACGTGCTCGGAC
CGCGGGAGCGGCACCGGGTCGTCGAGGAGTGGAACGCGACGGCGAAGGGGCTCGCCCCGGC
CACCCTGCCCGAGCTGTTCGAGCGGCACGTGCGGGAGCGCCCCGGCGCCGAGGCGGTGGTCG
CCGGCGACACGTCGCTGAGCTACGCGGAGCTGAACGCGCGCGCCAACCGGCTGGCCCGGCTG
CTGGTGGCGCGGGGCGCGGGCCCCGAGCGGCTGGTGGCGCTGGCGCTGCCCCGCTCGGCCGA
ACTGCCCGTCGCCGTCCTCGCGGTGGCCAAGGCGGGCGCCGCCTACCTGCCGCTGGACCCGG
CCCACCCGGCGGAGCGGATCGCGGGCACCCTCGACGACGCGGCACCGGTCGCGCTGCTGACC
ACGGCCGCGGTGGCCGCCGGCCTCCCGGACACGGACGTGCCCAGGCTGCTCCTTGACGAGGA
GCCCGCGGCCGGCGGTGGCGAGGACGCCGCCGACCTCACCGACGCCGACCGGCTCGCGCCGC
TGCTGCCGGGGCATCCCGCGTACGTCATCTACACCTCCGGCACCACCGGCCGCCCCAAGGGCG
TCACCGTGACGCACTCGGGCCTGCCCGCCCTGCTGGACATCTTCACCTCGCAGCTGGACGTCG
TACCCGGCAGCCGCGTCCTGCACCACCTCTCCCCCGCCTTCGACGGCGGGTTCTGGGAGCTGG
CGATGGGGCTGCTGACCGGTGCGGCCCTCGTCGTCGTGGAGCCCGGCACCGTGCCGGGGCCC
GCCCTGGCCGCGCTGGCGGTCCGCCACCGGGTCACCCACGCGGCGATCACCCCCGCGGTGCTC
CAGCTGATCCCCGAGGGCGCGCTGCCCGCCGGCACGACCCTGGTCGTCGCCGCCGAGACCTG
CCCGCCCGAACTGGTCGCCCGCTGGTCCGCGGGACGCCTGATGCGCAATTCCTACGGGCCGAC
GGAGACCACGGTCTGCGCCACCATGAGCGCCCCGCTGGCGGGCGCGGCCGTCCCGCCGATCG
GTAGGCCGATCGCCGACACCGCCGGGTACGTCCTGGACGACGCGCTCCAGCCGGTGCCGCCG
GGCGTGCCCGGGGAGCTGTACGTGCGCGGGCCCGGACTCGCCCGCGGCTACCTGGGCCGCCC
GTCGCTGACCGCGGGCCGCTTCGTGGCGTGCCCGTTCGGGCCGGCGGGCGGCGTGATGTACCG
CACCGGCGACCTGGTACGCCACCGCGCGGACGGCGACCTGGAGTACCTGGGCCGCACCGACA
CCCAGGTCAAGCTGCGCGGCATGCGCGTAGAGCCGGCCGAGATCGAGGCGGTGACGGCGGGC
CTGCCCGGCGTGGCGCAGGCGGCCGTGCTGGTCCGCGAGGACACGCCCGGCGACCGCCGCCT
GGTCGGCTACGTCGTGCCGGACGCCGGGGCGAGTGTGGACCCCGGCGCGCTGCGGCAGGCGC
TGCGCGGCTCGCTGCCCGAGTACATGGTGCCCGCCGCCCTCGTGGTCCTCGACGCGCTGCCGC
TCACCACCAACGGCAAGCTGGACCACCGTGCGCTGCCCGCCCCGGAGTACCGCACCGTCGAG
GGCCGCTCGCCGCGCACCCCGCGCGAGGAGGCCCTGTGCCGGCTCTTCGCCGAGGTCCTCGG
TCTGGAGCTGGTCGGTCTCGACGACGGCTTCTTCGACCTGGGCGGCCACTCGCTGCTGGCGAT
CCGGCTCGTGGAGCGGGTCCGTGCCGAGCTGGGCGAGGAGCTGGGCGTGCGCGACCTGTTCG
CCGCTCCCACGGTCGCCGACCTGGCGGTGCGGCTCGCGGCGCGCGGCGGCCGGGAGCCGATG
GAGCGGCTGCTCCCGCTGCGTGCCGCCGGTACCGCCCGGCCGGTGTTCTGCGTCCACCCCGGT
TCCGGCATGAGCTGGTGCTACTCCGGCCTGGTGCGCCACCTGCCGCCCGGTATCCCGGTGTAC
GGCCTCCAGGCCGCCGGACTCGACGGCGACGGGCCGCTGCCCGCGACGCTTCAGGAGATGGC
CGCCGAGTACGCGGATCTGGTCCGGCAGACGCAGCCCGAGGGCCCGTACCGGCTGCTGGGCT
GGTCGCTCGGCGGCAACGTGGCCTTCGCCATGGCCCGCGAACTCCGCGCCCGCGGGCAGGAG
GTGGAGCTGCTGGCCTTCCTGGACGCCTATCCGCGCCGGGCGGGCGCGGGCCCGGAGGCCCC
GCTGGCCGAGGTGTTCGCGCACAACCTGCGCGACGCCGGGTTCGACGTCGCCGAGGAGGAGC
TCACCGGCGGCCGCTTCCCCACGGCCCGCTACCGCGCGTTCCTGAACGCCGCCGGGGACCCGA
TGGGCCGCCTGGACGAGGCCGAACTGGCCGCGGTCCTGGAGGTGTTCATGAACAACGCGGCC
CTGATGCGCGGCCACACCCCCGGCACCTACGACGGCGACGTGCTGGTGCTGGCGGCCGAGCG
GGCGGACGGCGACAAGCTGGCCCGCCGCGGCGCCGAGTCGTGGCGCCCGCACGTGCGGGGC
CGGATCGAACGGGTCGGTGTCGACGCCGACCACCTCGGCCTCGTGCAGTCCGACGCCGCCCTC
GCCGTCATCGGCCGCGCGCTGGCCGGCCGCCTGGACCCGGCGACGGGCCACGCGGCTTCCGC
AGCAGTTCCCGAGACCGAAGGAGTCACCGCCATGAACCCCTCGCCCGAGCCCGCCCCGTCGC
CCGAGTCGCTCGATTCGACCGAGGTCGCCTGA
2.SCO5088编码合成蛋白actinorhodinpolyketide beta-ketoacyl synthasesubunitbeta,全长1224bp,具体核苷酸序列如下:
ATGAGCGTCCTCATCACCGGAGTCGGCGTCGTCGCACCCAACGGCCTCGGCCTGGCGCCGTAC
TGGTCGGCGGTGCTGGACGGCCGCCACGGCCTCGGCCCGGTCACCCGCTTCGACGTGAGCCG
GTACCCGGCGACCCTCGCCGGCCAGATCGACGACTTCCACGCGCCGGACCACATCCCGGGCCG
GTTGCTGCCGCAGACGGACCCCTCCACACGGCTCGCGCTGACCGCCGCGGACTGGGCCCTCC
AGGACGCGAAGGCGGACCCGGAGTCGCTGACCGACTACGACATGGGCGTGGTGACCGCCAAC
GCCTGCGGTGGGTTCGACTTCACCCACCGGGAGTTCCGCAAGCTGTGGTCCGAGGGCCCCAA
GTCGGTCAGCGTGTACGAGTCCTTCGCCTGGTTCTACGCGGTGAACACCGGCCAGATCTCCAT
CCGGCACGGTATGCGCGGTCCGAGCAGTGCCCTGGTCGCCGAGCAGGCCGGGGGCCTGGACG
CCCTGGGCCACGCCCGCCGTACGATCCGCCGCGGCACGCCGCTGGTGGTCTCCGGCGGGGTGG
ACTCGGCACTCGACCCGTGGGGCTGGGTCTCGCAGATCGCCAGTGGCCGGATCAGCACGGCC
ACCGATCCGGACCGGGCGTATCTGCCCTTCGACGAGCGGGCGGCCGGCTACGTTCCGGGCGAG
GGCGGGGCCATCCTCGTCCTGGAGGACAGCGCCGCGGCCGAGGCCCGCGGCCGGCACGACGC
GTACGGCGAACTCGCCGGCTGCGCCTCCACCTTCGACCCCGCCCCCGGATCCGGACGGCCCGC
GGGCCTGGAACGCGCGATCCGGCTGGCGCTGAACGACGCCGGGACCGGTCCCGAGGACGTCG
ACGTCGTGTTCGCCGACGGCGCGGGCGTGCCCGAGCTGGACGCCGCCGAGGCCCGGGCGATC
GGCCGGGTGTTCGGCCGCGAAGGGGTGCCCGTCACCGTCCCCAAGACGACGACGGGCCGCCT
CTACTCCGGCGGCGGCCCGCTCGACGTGGTCACCGCCCTGATGTCCCTGCGCGAAGGGGTGAT
CGCGCCGACCGCGGGCGTGACGTCCGTACCCCGGGAGTACGGCATCGACCTGGTGCTCGGCG
AACCCCGCAGCACGGCCCCGCGCACCGCACTGGTCCTGGCCAGGGGCCGCTGGGGCTTCAAC
TCGGCGGCGGTCCTGCGCCGTTTCGCACCGACCCCGTAA
3.SCO5892编码合成蛋白polyketide synthase RedL,全长6894bp,具体核苷酸序列如下:
ATGGCCCGGAACCGCAGGGAGAAGAACGTGAGCCACGGGCCGACGGACCCGCTGCACGAGC
ACTGGCAGGGACCCGAACCGGAGACGGTCACGGCCCTGCTGCGGCACCGGGCGGAACACAC
CCCCGACGCCCTCGCCCACCGCTTCCTCACCGACACCGACGGTACGTCCGTCGCCTGGACCTA
CCGCGAACTCGACCTGCACGCCCGCGAGGTGGCCACCCACCTGCGGCACGCCGGGGTGGGGC
AGGGCCCGGTGCTGCTGCTGCACCCGCCGGGCCTCGACTACCTCGCCGCGTTCTTCGGCTGCC
TGTACGCCGGCGCCGTGGCCGTCCCCGCCTACCCGCCGGACAACGCCCGGTTCGGCCAGACCG
TGCCCCGGCTCGCGGCCATCGCCCGGGACTGCGCCGCCACCCACGCCCTCACCACCCGCCGGG
TCCGCGAGACCGTCGCCGCCGACGGCACCGGCCGGGTCGGCACCGAACTCGACGGCGTGCGC
TGGCTGGTGACCGAGGACCTCTACACCGGCGAGTCCACCACCGCCTGGGAGGACCCCGGAGC
CACGGCGCGGTCGCTGGCCTTCCTCCAGTACACCTCGGGTTCCACCGCCGCGCCCAAGGGCGT
GATGGTCGAACACGGCAACCTGGTGCGCAACCTGCGCTCCATCCACCTCAGGCTCGGGCACG
ACGCGGACTCCGGCATGGTCTCCTGGCTGCCGCCGTACCACGACATGGGTCTCATCGGCGGCA
TCCTCACCCCGGTGTACGGCGGCTTCCCCGCGCACCTCATGGCGCCGATGACCTTCGTGCAGC
GCCCGCTGCTGTGGCTGGAGACCCTCTCCCGCACCGGCGCCTCCACCAGCGTGGCACCCAACT
TCGGCTTCGAGCAGTGCCTGCGCCGGATCACCCCCGCACAGCGGGCCGGCCTGGACCTGAGC
CGCTGGCGCCTGGCGCTGAACGGTGCCGAGCCGATCCGCCCCGACACCCTCGACCGGTTCGCC
GAGTACTTCGCCCCGGCCGGCTTCGACCGCACCGCGCTGCTGCCCTGCTACGGCCTCGCCGAG
GCGACCCTCATGGTCACCGGCGTGCGGCCGGCCGACCCGCCGGTGGTGGAGTCCTTCGACGC
GGCGGCCCTGGAGGCGGGCACCGCGCGTCCCGCGGACCCCGGCGGCGTGCGCACCACCCGCG
TCGTCGGCTGCGGAGCACCCGTCGCCGACGTCGAGGTCGCGGTGGTCGACGCCGCCACCGGC
CGCCGGGTCCCCGACGGCACCGTCGCCGAGATCCGGGTCAGTGGTCCCGGCGTCGCCCGCGG
CTACTGGGGACGCCCCGAGGCCGCCGCCGAGGTCTTCGGCACCCGGATCGACGGCGAACCGG
GCAACGCCTGGCTGCGCACCGGCGACGTCGGGTTCCGCCACGACGGCCAGCTCTACGTGGTC
GGCAGGACCAAGGACGTGATCATCGTCCAGGGCCGCAACATCCACCCCCAGGACGTGGAACA
GACGGCCGAACGGGTCGGCGCCGGACTGCGCGCCGGGTACGGCGCCGCGTTCGCCGTCCCCA
CCGCCGACGGCGAACAGCTCGCCCTCGCCTACGAGATCGGCGGCCCCGGCGCCGGGGATCCG
CACGCCCTGCTCGCCCGGCTGCGCACCGCCATCGCCGACGAGCACCAGGTGACACCGCACAC
CGTCGTGCTGCTCAAACGCTCCTCGGTGCCGCGCACCACCAGCGGCAAGATCCAGCGCGGCG
CCTGCCGGCAGCGCCTGCTCGACCTGGAACTGCCCGTGGTGGCCGCGAGCGTCGTCCAGGAC
GGTGCCCTGCCGGACACCCCCGCCCCGCACGCCCCCGCCTCCCCGGACCGTACGGCCGCCCTC
GTCGCCGACGCCCTCGACCGCGCCGCGACGGACGCGCCCGACGGCACCTCGCGCTACCCGCG
GCTGCTGACCGCCGTACGGACGCTGTCCGAGCGCCTCGGCGTCGCCCTCGACGTCGGTGAACT
CCTGTCCGGCCCCGAGCCGGACGGCGCGCCGTCCCTCACCGGGCTGTGCCGCGCCGCGCTCG
CCGACGCGGCGCCCGCGCCCACGGCCGCGCCGGAGCGGACCCGGACCCAGGCCGTGGACTG
GCTGCGGCGGGCCGTCGCCGCCCGGCTGGGCCTGCGCCCCTCGGTGATCGACACGGCGGTCC
CCTTCACCTCACTGGGCCTGGACTCCAAGCAGGCCGTGGCCCTCACCGCCCCGTTCGGCGACT
GGCTCGGCACCCCCGTGACCCCGGCCACCCTCTACGACCACCCGACGGTCGAGTCCCTGGCG
GCCCACGTCACCGGACCAGAGGCGGACGGCACCCCGCACCCGCACCCGCGGCGTCCCGCTCC
CGAGCCCGCCCGCGGGCGCGCCGACGAGCCCGTCGCCGTGATCGGCATCGGCTGCCGGTTCCC
CGGCGCACCCGACCCCGACAGCTACTGGCGGCTGCTGCGCGACGGCCGCGACGCCGTGGGCG
AGGCACCCGCCGACCGCCACGGCGACACCGGCGGCCGGCCGGGCGGCTTCCTGGAGCACGTG
TACGACTTCGACGCCCGCTTCTTCGGCGTCTCCGCCCGCGAGACCGAACGCATGGACCCCCAG
CAGCGGCTGCTCCTGGAGGTCGCCTGGCAGACCTTCGAGGACGCGGGCATCGCGCCCTCCTCC
CTGGCCGGCACGGACACCGGTGTCTTCGTCGGCATCAGCGGGCACGACTACGCCGATCTCCAG
ATGCCGCACCCCGACGTCGTCGACATGTACTCCGCCACCGGCAACGCCCAGTCCGTGGCGGCC
GGCCGGCTCTCGTACTTCTTCGACCTGACCGGGCCCAGCCTCGCCCTCGACACCGCCTGCTCC
TCCTCGCTCGCCGCCGTCCACACGGCGCTGCGCAGCCTGCGGGACGGCGAGTGCGGCGTCGC
CCTCGCGGGCGGCGTCAACCTGATGCTGACGCCCGGGCTGTCGGAGGCCCTCGCCCGGGGCG
GCATGCTCGGCCCCGGCGGACGCTGCCGCACCTTCGACGACGGCGCCGACGGCTACGTCCGC
GGCGAGGGCGCGGGCCTGGTGTGCCTCAAGCCGCTGTCCGCCGCGCTGGCCGACGGGGACCG
CGTGCACGCCGTGCTCACCGGCTCCGCGCTCGGTCACGGCGGCCGGGCCAACGGCCTGACCG
CCCCCCGCAGCACCGCCCAGCGTGCCGTGATGACCGGGGCGCTGGAGCGGGCCGGTGTGCAG
CCCGGACAGATCGACTACGTCGAGGCACACGGCACCGGCACCGCTCTCGGCGACCCCATCGA
GTGGGAGGCCCTGGCCGGCGTCTACGGCCGCGGCCGCCCCGCCGACGCGCCCTGCCCGGTCG
GCTCGGTGAAGACCGCCATCGGCCACCTGGAGGCGGCCGCCGGCATCGCCGGGCTGATCAAG
GCCGTGCTCGTGGTGCGGGAGCGGGAGGTACCGCCGCTGCTGCACCTGGCCACCCCCAACCG
GCACCTGGACTGGACGGGTTCGGGACTGACCGTGCCCACCACCCGCCGGGCCCTGCCCGCCG
GGGGCACCCTGCGCGCGGGCGTCAGCTCGTTCGGCTTCGGCGGCACCAACGCCCATGTGATCG
TCGAGTCCGCGCCCGCACCCCCGGCGCCCGCCGGCGAGTCCGGCGCCCCCGACGGCACGGTG
CACGCCCTGTCCCTGTCCGCGCACACCGAGTCCGCCCTGGCCACCCTGGCCGGCCGCTACCGC
ACCCACCTCGCGGCCCACCCCGGCGTGCCGCTCGCCGCCCTGTGCCACTCCGCCCGCACCGGC
CGTTCCCGGCTGCCGCACCGGGCGGTCCTCACCGCAGCGACCCGGGACGAACTGGACACCGC
ACTGGAGGCGCTGATCCGCCGGGAGCCCGCCGTGGACGTGGCCCGCGGCGGACCCGCGGACG
GCACCGCGCCCACCGTCGCCTTCCTCTTCGGCGGCCAGGGCACCCAGTACCCGGGCATGGGCC
GCGACCTCTACGACGCCCACCCCGTCCTCGCCCGCACCCTGCGGCGCGCCGACGCGGTGCTGC
GCGGCCTGGGCGAGATCCCGCTGCTCGACCTGCTCTTCGACCCGGACCGCGCCGAGGACCTG
GCCCGCACCCGGTACTGCCAGCCCGCCCTGGTCGCCCTCGAAGTCGCCCTCGCCGACCTGTGG
ACGAGCTGCGGAGTACGCCCGGCGGCCGTGCTCGGGCACAGCGTCGGCGCCCTCGCCGCCGC
CTGCGTCGCCGGTGTCCTCTCCCTGGAGGACGCCCTCACGCTGGCGGTCGTCCGCGGGCGTGC
CATGGACGAACAGCCCGGCGAGGGCGCGATGATCGCCTGCGTCGGCGATCCGGAGACGGTGC
GCGAGATCGCCGCCGGCCACGACCGGGTGGCCCTCGCCGCCGTCAACGCGCCCCGCAACCTG
GTGTTCTCCGGCGCGGCCGACCGGATCGCGGCACTCCGCGGCGACCTGGAACGCAAGGGGGT
CACCGTCCGGCCGCTCGCCGTCTCGCACGCCTTCCACTCACCGCTGATGGCGGGCGCCGCCGA
GCCGCTGCTGGAGGCCGCGCGCGCCGTCGAGTTCCAGCCCCCGGCCATCCCGTGGATCTCCGA
CGCCACCGGCGAACCCGTGACCCGGGTCGACGCCGAGTACTGGGTCCGCCACCTGCTCGGCA
CCGTGCGCTTCGCCGACGGCTTCGCCCGCCTGCGCGCGGGGAACGCCGAAGGCGGTCTGGGC
TGCGACGCGTTCGTCGAGATCGGCCCGCACCCCACCCTGCTGAACCTCGGCCGGTCCGCCGTC
GCCGACGGCCCCGAGCCCGACGGACGCCGCACGCTGTGGCTGCCCTCGCTGCGCCGCGGCGC
CCCCGGCCGCCGCACCCTGCTGCGGTCGCTGGGCGGCTACCAGTGCGCGGGCGGAGACATCG
ACTGGTCCGCCCTGGACGACGAACACCCGCCCGCCCGGGTCCCGTTGCCGCACACGCCCTTCG
AACGCCGGACCTACCGGTTCCAGGCCCCCACTCCCGATCAGGAAGGCGATCCCATGCCGTCCC
AGCCCACGGTGCCGGGCACCGCCCCGACGGGCCCCGCGGGCGAGGTCTCCCGCACCGTGCTC
GACACCGTCGCCCGCGTCTGCGGATTCCCCGTGGCCGACCTGCCGGTGCACGCCAGGCTCGGC
CTCGAGCTGGGATTCGACTCCCTGATGCGGACCGACCTCCAGCGCCGTCTCACCGACCGCTTC
CCCCAACGGATGCACACGCTCCAGGACCTGCCGGAGGACCCCACGGTGCAGGACATCATCGG
CCGGCTCGCCGCCGGCGACGACCGCGACGGCGACCCCGCCGGCGGCGCCCACACCGCCCGCC
CCGCCCTCGCCGGCACCGGCCCGGCCGCGCCGCCGGCCCACCCCGCGGCCGTACCGGCGGCG
CCGCCCGTCCCGGCCCCGCAGTACGCCCCGTACGGAGCCTTCCCGGCCCCGCCCCACGGCTAC
GCCGTCGCCGCCCCGGTCCCGCAGCCCGGAGTGCCGTACCCGTACCCGTACGGCTACCCGCCG
CACGGTGCTCCCGGGTACGCCTACTACGTCCCCCAGCCCTACCCCCCGCAGGCGTTCCCGCCC
CCGGCACCCCCCTCCCCGCTGCAGGCGGCCCCCGCCCCCGTACCCGCCGCAGCCCCCGCCCCC
GTTCCCGACCCGGTTCCGGCGCCCGCCGCCCCCGTACGTCAGGAGCGCGCCTTCGAGGACTGG
GCGGAGTACGCGGAACTCCAGGGCAGACTCCGCCAGACCCGGACCAGCGGGTCCAACCCCTA
CGGCCGTACGCACGAGGGCTTCAACTCCGCGCTCGCCACGGTCGACGGCACCAAGGTCGTCA
ACTTCGCGGCCTTCAACTACCTGGCGCTCTCCCACCACCCCCGCGTGCGCCAGGCGGCCAAGG
ACGCCGTGGACCGCTACGGCACCTCCGCCTCCGCCACCCCGCTGCTGTTCGGCGAGACCCCGC
TGCACCACGAACTCGAAGCCGAGATAGCACGGTTCATCGGTACCGAGGCGGCCATCGTGTTCG
CCGGCGGCCACGCCACCAACGTCGCCACCGTCGGCCACCTGTTCGGCCCGGAGGACCTGGTG
GTGCACGACGCCTGGATCCACGACAGCACCGTGCGCGGCTGCATCCTCTCGGGCGCCCGGCGC
CGGTCCTTCCCGCACAACGACTGGGCCGCCCTCGACCGCATCCTGACCGGAGCACGGGCCTCG
CACCGCAGGGCGCTCGTCGTCATCGAGGGCGCCTACAGCCAGGACGGCGACATCCCCGACCT
GCCGCGCTTCATCGAGGTCAAGAAGCGCCACGGCGCCATGCTGATGATCGACGAGGCGCACTC
CATCGGCGTCCTGGGCCGCACCGGCCGCGGCGTGGGCGAGCACTACGGCACCGACCCGGCCG
ACGTCGACCTGTGGATGGGCACCCTCAGCAAGGCGCTGGGCAGCCTCGGCGGGTACATCGCC
GCCCGCCGCCCGATCATCGAGTACCTCAAGTTCACCGCGCCGCTGCACATCTTCTCCACCGGCA
TCTCCCCGGCGAACGCCGCCGCCGCCCTGGAGGCGCTGCGGGTCGTCCAGGACGAGCCCGAG
CGGGTCGCCCGGGTGCAGGAGCTGGCCGAGTTCTTCCGCGCAGGCGCCCGCGCCCGCGGCCT
CGACGTCGGCGTCTCCCGCCTCTCCGCGGTGATCCCGGTGATCACCGGTGACTGGGAGAAGAC
GATGGCGCTCTCCAACTCGCTGCTGGAGCGCGGCGTCAACGTCATGCCGATCGGCTACCCCGC
CGTCCCGCGCGACCAGTGCCGGCTCCGCTTCTTCATCAACGCGGACCACAGCGAAGCAGACCT
CGAACACTCCCTCGACCTCCTGGTGTGA
4.SCO6274编码合成蛋白type I polyketide synthase,全长10731bp,具体核苷酸序列如下:
GTGGCCAACGAGCAAGAACTCCGCGAGTACCTCAAGCGGGCGATCGCCGATGCGCGCGACGC
GCGCAAGCGCCTGCGCGAAGCCGAGGACAAGGCCCACGAGCCCATTGCGATCGTCGGCATGG
CCTGCCGCTACCCCGGGGGCGTCGCCACGCCGGAGGACCTGTGGCGGCTGGTCGACGACGGC
GTGGACGCGGTCGGCGGCTTCCCCGTCAACCGCGGGTGGGACCTGGAGCGGCTGTACGACCC
GGACCCGGACGCCGCGGGCACCTCGTACACGACCGAGGGCGGCTTCCTCCACCAGGCCGACG
AGTTCGACCCGGAGTTCTTCGGGATGTCGCCGCGCGAGGCACTCGCCGTGGACCCGCAGCAG
CGGCTGCTGCTGGAGACCACCTGGGAGGCGTTCGAGCGGGCGGGCCTCGATCCGGAGTCGTT
GCGCGGCAGCCGCACCGGCGTCTTCACGGGCTTGATGTACAACGACTACGGCTCCCGCCCGAA
CCTCCCCGCCGAGGGGAACGAGGGCTATCTGTTCAGCGGCAGCGCGGGCAGCATCGCCTCGG
GCCGGCTGGCGTACACCTTCGGGCTGGAGGGCCCGACGGTGACGGTGGACACGGCGTGCTCG
TCGTCGCTGGTCGCCCTGCACATGGCAGCGAACGCGCTGCGTTCCGGTGAGTGCGACCTCGCG
CTCGCGGGCGGCGCGGCCGTGATGTCCACGCCGACCGCCTTCGTCGAGTTCTCCAGGCTGCGC
GGACTGGCTCCGGACGGTCGCTGCAAGTCCTTCTCCGAGCACGCGGACGGCACCGGCTGGGC
GGAGGGGGTCGGGCTGCTGCTCGTCGAGCGGCTCTCCGACGCCCGCCGCAACGGCCACCAGG
TACTGGCCGTCGTCCGCGGCTCCGCCGTCAACCAGGACGGCGCCTCCAACGGCCTCACCGCGC
CCAACGGCCCCTCCCAGGAACGGGTCATCCGCCAGGCCCTGGCCAACGCGGACCTGACCCCC
GCCGACGTCGACGCCGTCGAGGCACACGGCACCGGCACCACCCTCGGCGACCCGATCGAGGC
CCAGGCACTGCTGGCGACGTACGGCCAGGACCGGGCGGAGGGCCGGCCGCTGTACCTGGGCT
CGCTGAAGTCCAACATCGGACACAGCCAGGCCGCCGCCGGAGTCGGCGGGATCATCAAGATG
GTGCAGGCGATCCAGCACGGTGTGCTGCCCGCGACCCTGCACTCCGAGGAGCGTTCGAGCCG
GGTCGACTGGGAGGCGGGCGCGGTCGAGTTGCTGACCGAGGCCCGGCCCTGGCCGGAGACCG
GCGCGCCGCGCCGGGCCGGAGTGTCGTCGTTCGGCTTCGGCGGCACCAACGCGCACGTGGTC
ATCGAGCAGGCCGTGGAGCCGACGGACGAGGATGCGCGGGACAGGGACGAGCCCACGTCCG
AGGCTCCGGCGCAGGCGGACGTCTCAGCGCCGCCGGTGCTGCCGTGGGTGCTGTCCGGGCGC
ACCCGGGAGGCCGTGGCCGGTCAGGCGCGCAGGCTGCTGTCGTACGCGACGGAGCACGAAGA
CGTCGAGCCCGCCCATGTCGCCCGGTCACTGGTCCGCGACCGGTCGGTGTTCGACCACCGTGC
GGTGGTGGTGGGGTCGGACCGGGAGGAACTGCTCACCGGACTACGAGCCCTCGCCGAGGGCC
GGACCACCCCCACCGGCGTCGTCCAGGACACCAGGCAAACCGGCAAAACCGCGTTCCTCTTC
ACCGGCCAGGGCGCCCAACGCACCGGCATGGGCATGGACCTCTACCGCACCTTCCCCGCCTAC
GCCCACGCCTTCGACACCATCACCGCCCAACTCGACCCCCACCTCGACCAACCCCTCCACCAC
ACCATCACCACCGGCCACCACCTCCACCACACCGGCAACACCCAACCCGCCCTCTTCGCCACC
GAAGTCGCCCTCTACCGACTCCTCGAAACCTGGGGCATCACCCCCGACTACCTCGCCGGACAC
TCCATCGGCGAACTCACCGCCGCCCACATCTCCGGCATCCTCACCCTCCAAGACGCCTGCACC
CTCGTCACCGCACGCGCCCGCCTCATGCAAAACCTCCCCACCCACGGCACCATGATCGCCCTC
CAAGCCACCGAACAAGAAATACTCCCCCACCTCACCGGACACGAACACCACCTCACCATCGC
CGCCATCAACAGCCCCACCAGCCTCGTCATCTCCGGCAACCAAACCGCCGCCCAACACATAGC
CACCCAACTCCAGGCCCAAGGACGCAAAACCAAAACCCTCACCGTCTCCCACGCCTTCCACT
CCCCCCACATGGACGGCATGCTCCACGACTTCCACCACACCGCCACCCAACTCACCTACCACG
AACCCACCATCCCCATCGTCTCCACCCTCACCGGAAACCTCGCCACCCACAACGACCTCCGCA
CCCCCACCTACTGGACCGACCAACTCCGCAACACCGTCCGCTACACCCAAGCCATCCACACCC
TCCACACCGCCGGCGTCACCACCTACACCGAAATCGGCCCCGACGCCACCCTCACCCCCCTCA
CCGCGAGCACGGTCGAGGACTCCGACGGCGTGGCGGCCATCGCCACCCTGCGCGCCGGCCGC
CCCGAGCCGCAGCAGGTCCTCGCGGCGGTCGGCGAGTTGCACGCACGCGGCCGACGCGTCGA
CTGGGAAGCGTTCTTCGGCGGCCGGCCCGGCCAGTCCGTGTCCGTGGACCTGCCGACGTACGC
CTTCCAGCGCGACCGGTACTGGCTAGACGTCACGGAGGCCGCCGCCGACGCCGCCGGCCTCG
GGCTGACCCCGACGGACCACCCGATCCTCGGTGCCACGCTCGATCTGGCCGACGGCGAGCAG
ACCCTCTTCACCAGTCGGCTGTCCCTCCGCACACATCCCTGGCTGGCCGACCACACCGTGGCC
GGCACCACTTTGCTGCCGGGTACGGGCTTCGTGGAACTGGCCGCGCTGGCGGGTGAGCGGTT
GGGCTGCCCGCGGGTCGAGGAGTTGACGCTGTCGGCGCCGCTGGTCCTGCCGGAGCGGGACG
GCGTGCGGATACAGCTCGCGGTCGGCGAGGCCGACGGCGCGGGCCGCCGGGTGGTCGACGTG
TACGCGCGTCCCGACGGCGGCGACGAGACACCCGGCGCCGTTGCCAAGGCCCGCCCGTGGAC
CGCGCTCGCCAAGGGCGTGCTGGCGCCCGCCACCGGAACCGGCACAGCGGCGGACGGTCTGC
CGGTCTGGCCGCCGACGGGCGCGAGTGAGGTGCCGCTCGACGGCGTCTACGACCGACTGGCG
GCCCAGGAGTACGCGTACGGTCCGGCTTTCCAGGGTCTGCGGCGGGTATGGCGCGGCGACGG
CGAGATCTACGCCGAGGTCGCCCTGCCCGAGGAGCAGCGCGCCGACGCGGGACGCTACATCC
TGCACCCGGCCCTGCTCGACGCCGCACTGCACCCCCTGCTGCCCGGCGTCGCCGACCAGGACC
GCGCGGCGCTGCTGCCCTTCGCCTGGTCCGGCGTCACCATCCACGCCACAGGCGCCTCCGTAC
TGCGCGTGCGGCTGACACTCACCGGGTCCGACGTCGCGGCGTTGACGGTGGCGGACGCAACC
GGCGCCCCGGTCGCCTCCGTCGAATCCCTGCTGCTGCGCCCCCTGTCCAAGGACGCACTGCGC
CAAGCCGCCTCCACCGCCCGCGACGGACTCTTCCGCATCGCCTGGAACACACTGCCCACCACC
GACACCCTCACCGACACCACCGGCTGGGCCGTCGTCGGCGACCTGACGGTCGACGGAGCGAG
CAGGCACGCCGACCTCGGCGCCGTGGCGAACGCCGATTCCGTCCCGGACACCGTCCTGTACGC
TCCCCCGGTCCCGGACGGTGACGTGCCGGAAGCGGCCCACACCGCACTGCGCGACGCCCTCA
GCACCACGCAGTCCTGGCTGGCCGACGACCGCACCACCGACACCACCCTCGTCGTCGTCACC
CGGGGCGCCCTCGCCACCCGCGAAGGACAGCACACCGACCCCGTACAAGCCGGCCTCTGGGG
CCTGCTCCGCGTCGCCCAGACCGAGAACCCCGGCCGCATCGCCCTCCTCGACACCGACACCGA
CACCATCCCCACCACCGCCCTGACCACCGACGAACCCCAACTCGCCCTCCGCGACAACACTTT
CCACACCCCTCGCCTGGCCCGCACCACCGACCAGCCCACCGACCAGCCCACCGACCGGCCCC
GCTGGGACCGCGGCACCGTCCTCATCACCGGCGCCACCGGCGCCCTCGGCACCGTCCTCGCCC
GCCACCTCGTCACCCAGCACGGCACCCGCCACCTTCTCCTCCTCAGCCGACGCGGCACCAACG
CCCCCGGCGCACAAGAACTCCAGCATGAACTCACCGCCCTCGGCGCCACCGTCACCATCACCG
CCTGCGACGCCGCCGACCGCCACGCCCTCACCTCCGTACTCGACGACATCCCCGCCGAACACC
CCCTCACCGCCGTCGTCCACACCGCCGGCGTCCTCGACGACGGTCTGCTCTCCTCGCTCACCG
AGGAGCGGCTCGCCGATGTGCTGCGTCCGAAGGTCGACGCGGCGTGGAACCTGCACGAACTG
ACCAAGGACCACGACCTGGAGGCATTCGTCCTCTACTCCTCCGTCGCCGGCCTCATCGGCAAC
GCCGGCCAGGCCAACTACGCCGCCGGAAACACCTTCCTGGACGCCCTCGCCCAGCACCGCAG
GGCACTCGGACTGCCCGCCGCCTCCCTCGCCTGGGGCCTGTGGGCCCAGGCCAGCACCATCAG
CGGACAACTCGACCAGACCGACCTGAAGCGCATGGAGCGCACCGGGCTGCTGCCGCTGTCGT
CCGCCGACGCGATGGACCTGTTCGACGCGGCCCCGGCCACCGGCGAAGCGGTCCTCGCGGCC
ACCCGGCTGGACCTCGGCGCACTGCGCAAGCAGAGCGACGGCCCGCACGTCCTCTTCCGCGG
CCTCCTCCCGGCCGCCCCGCGCCGGGCGGCCGCCGCGCAGGCCGACGGCGGTTCGTCGCTGG
AGCAGCGGCTGGCTCCGCTGTCCGCGGCCGAGCGCGAGCGGGCACTCACGGATCTCGTGCGC
GTCCAGGTCGCGGCGGTGCTCGGGCACTCTGACCCCGGCGCGATCGAGTCCGGCCGGGCCTTC
CAGGAGCTGGGCTTCGACTCACTGACAGCCGTCGAACTTCGCAACCAGCTGAGCACCGCGAG
CGGACTGCGCCTGCCCACCACCCTCGTCTTCGACCACCCCTCCCCCGCCGCTCTCGCCGCCCA
CCTCTCGGCGGAGCTGTTCGGCGAGCAGGAGTCGAGCGGGCCGGCCGTGGTGGCCGCCGGCT
CGGCGTCCCTGGAGCCGGTCGCGATCGTCGGCATGGCCTGCCGCTACCCGGGCGGGGTGTCCT
CGCCGGAGGAGCTGTGGCGGCTGGTGGCGGACGGGGTGGACGCCGTCACCGAGTTCCCCGTC
AACCGGGGCTGGGACCTGGAGAACCTGTACCACCCTGACCCTGAGCACGTGGGGACGACGTA
TGCCCGGGGCGGCGGTTTCCTGCACGACGCCGACCTGTTCGACCCGGAGTTCTTCGGCATGTC
GCCGCGCGAGGCGCTCGCCACCGATCCGCAGCAGCGGCTGCTGCTGGAGACCGCCTGGGAGA
CGGTCGAGAACGCGGGCATCGTCCCCGCGTCGCTGCGCGGCAGCCGCACCGGTGTGTTCACC
GGCGTGATGTACCACGACTACGGTTCCCAGCTGGACACGGTCCCCGAGGACCTGGAGGGCTAT
CTCGCCAGCGGCAACGCCGGCAGTGTGGCTTCGGGCCGGGTCTCGTACAACCTGGGCCTGGA
GGGACCGGCGGTCACCGTCGACACGGCGTGCTCCTCGTCGCTGGTCGCCCTGCACATGGCGGC
GAACGCGCTGCGTTCCGGTGAGTGCGACCTCGCGCTCGCGGGCGGGGTCACCGTCATGTCGA
GCCCGACCTCCTACGTGGAGTTCTCGCGGCAGCGGGGCCTGTCGCCGGACGGCCGCTGCAAG
CCGTTCGCGGCCGCCGCGGACGGCACCGGCTGGAGCGAGGGCGTCGGCCTGCTGCTCGTCGA
GCGGCTCTCCGACGCCCGCCGCAACGGCCACCAGGTACTGGCCGTCGTCCGCGGCTCCGCCGT
CAACCAGGACGGCGCCTCCAACGGCCTCACCGCACCCAACGGCCCCTCCCAGGAACGGGTCA
TCCGCCAGGCCCTGGCCAACGCGGACCTGACCCCCGCCGACATCGACGCCGTCGAGGCACAC
GGCACCGGCACCACCCTCGGCGACCCCATCGAGGCCCAGGCACTGCTGGCGACGTACGGCCA
GGACCGGCCCGACGGACGCCCCCTGTACCTCGGCTCACTCAAGTCCAACATCGGACACAGCC
AGGCCGCCGCCGGAGTCGGCAGCGTCATCAAGATGATCGAGGCGATGCGCCACGAGGTGCTG
CCCAGGACCCTGCACGTCGACGCGCCGTCCCCGCACGTCGACTGGGAGACGGGCGCGGTCGA
ACTGCTCACCGAGGCCCGGGAGTGGGAGACGTACGGCCGTCCGCGCCGCGCCGCCGTCTCGT
CCTTCGGCATCAGCGGCACCAACGCCCACGTGGTGATCGAGCAAGCCGAGCAGCCGGAGCCG
ACGGAGCCGAACGGTCGGCCCGAGCAGCCGGAGCCGGTGTCCGCCGCCCCGCTGCCGGCCGT
CCCGTGGGTGCTGTCGGGCCGGACCCCGGAGGCCGTGACCGGACAGGCGCAGCGGCTGCTGT
CGTTCGTCGAGTCACGCCCCGAAGCGGCCCCGGCGGACATCGGGCTCTCGCTGGCGGTCAGC
CGGTCGCAGTTCGACCACCGGGCGACGGTCGTCGGCGCGGACCGCGAGGAACTGCTGGCCGG
GCTGCGGACGCTGGCTTCCGGTGCTACGACGCAGACAGCGGTGCGGGACACCAGACAAACCG
GGAAGACCGCGTTCCTCTTCACCGGCCAGGGCGCCCAACGCACCGGCATGGGCATGGACCTCT
ACCGCACCTTCCCCGCCTACGCCCACGCCTTCGACACCATCACCGCCCAACTCGACCCCCACC
TCGACCAACCCCTCCACCACACCATCACCACCGGCCACCACCTCCACCACACCGGCAACACCC
AACCCGCCCTCTTCGCCACCGAAGTCGCCCTCTACCGACTCCTCGAAACCTGGGGCATCACCC
CCGACTACCTCGCCGGACACTCCATCGGCGAACTCACCGCCGCCCACATCTCCGGCATCCTCA
CCCTCCAAGACGCCTGCACCCTCGTCACCGCACGCGCCCGCCTCATGCAAAACCTCCCCACCC
ACGGCACCATGATCGCCCTCCAAGCCACCGAACAAGAAATACTCCCCCACCTCACCGGACACG
AACACCACCTCACCATCGCCGCCATCAACAGCCCCACCAGCCTCGTCATCTCCGGCAACCAAA
CCGCCGCCCAACACATAGCCACCCAACTCCAGGCCCAAGGACGCAAAACCAAAACCCTCACC
GTCTCCCACGCCTTCCACTCCCCCCACATGGACGGCATGCTCCACGACTTCCACCACACCGCC
ACCCAACTCACCTACCACGAACCCACCATCCCCATCGTCTCCACCCTCACCGGAAACCTCGCC
ACCCACAACGACCTCCGCACCCCCACCTACTGGACCGACCAACTCCGCAACACCGTCCGCTAC
ACCCAAGCCATCCACACCCTCCACACCGCCGGCGTCACCACCTACACCGAAATCGGCCCCGAC
GCCACCCTCACCCCCCTCACCCAGCAAACCCTCGACACCACCACCGCCATCCCCCTCCTACGC
CACAACCAACCCGAAACACACACCCTCCTCACCGCCATCAGCCGCCTCCACAACCGCGGCATC
CCCATCAACTGGAAAAACCTCTTCACCACCACCCCCGCCAAACACACCCCCCTACCCACCTAC
GCCTTCCAACACGAACGCTTCTGGCTGAAGGGCTCGGCGCGGTCCGGCGACGCGGGCCACGC
GCTCAGCCCGACGGGCCATCCGCTGCTGGGCGCCGCGGTCACCGTGGCCGGCGCCGATCAGG
TGCTGTTCACCAGCCGGCTGTCGGCGCACGGTCATCCGTGGCTGGCCGAGCAGGCGGTGTTCG
ACGCGCCCGTACTGCCCGTGTCCGCCCTGGTGGAGCTGGCCCTCCGGGCGGGCGACGAGCTG
GGGGTCCCGGGCGGGCTCGCGGAGCTGGACCTGCGCACGCCAGTCGTTCTGCCCGAACAGGG
CTTCATGCAGCTCCAGTTGGGCGTGGGGCCGGCGGACGAGGCCGGACGGCGGCCGTTCACCG
TGCACAGCAGGGTGGACGACGCGGACGCTCCTTGGAAGGTGCACGCCCATGGTGCCTTCGAG
TCCGCGGACGGCGGTGCGCCGCTTCCCGGTCCGTCGTCGGACGCGGATGCGACCGACGTGCG
GCTGGCCGAGGAACTGCTGCCGGACGCCGCCCGCTACGAGCTGCATCCGGCCCTGCTGGAGG
CGGCGGTCATGGCGGCCGCGGGGACCGGGGAGGCCGGCACCACGCCGGTACCGGCCCACTGG
CGGGGCGTGCGGCTGCACGCGTCGGGTGCCACCGCCGTACAGGTCCGGGTCGGCGCCCCGGA
CGAGCACGGCGTCAGCGTGCTGCTGTCCGATGCCGAAGGGCAGCCGGTCGCCACGGTCGAGA
GGCTGACCTTCCGGAATGTTCCGGACGCCGAGTTCGCGGTGGACTCCGAGGGCAGCCGGCCG
CTGTTCCGGGTCGACTGGGACCGGACCGCGCTTCCGGAGCCCGCTTCCCCGGTGCGCTGGGGC
GTCCTTGGTGACGGCGTCCGGTTCGACGGACCGGCCGACGGCATCATGGCGTTCGAGGACGTG
GCGGCCGTCGGCAAGGCGGCGGAGGCAGCCGGGACCGATGCCCCGGTGGACGCGGTGCTGG
CACGGATCACGCCCGAGGGGACCGGCGGTGCGCCGACGGCCGCGCACGCGGCTGCGCGCGGT
GCGCTGAACCTCGTACAGGACTGGCTGGCCGACGACCGGCTCGCCACCACCCGGCTCGTGGT
GGCGACACGAGGGGCGGTGCACGCCGGTGGGGAGCAAGACGCGCCGGACGTCGGTGCGGCG
ACCGTCTGGGGGCTGCTGCGCTCGGCGCAGGCGGAGGCCCCCGACCGGATCGTGCTCGTCGA
CCTGGACCACGCCGACGAGGGCACGGAGCCGTCGGTGTCGTCTGCCCCGCTGTCCGCGCTCG
TCGCGTCGGGCGAACCCCAGGCGGCGCTGCGTGGCGGCGAGACCCTGCTGCCGAGACTGCGT
CGGCTGCCGGCAAAGGCGTCCGACGGCTCCCCGGCCGAACCGGTCTGGGACCCGGAGGGCAC
CGTGGTCGTCACCGGCGGTACGGGCGCACTCGGCAGTCTGTTCGCCCGGCATCTGGTCGCCCG
GCACGGCGTACGCCACCTGCTGCTGCTCAGCCGGCGGGGCGAGGACGCGCCGGGGGCCGGCG
AGCTGATGGCCGACCTCGCCGCGCTCGGGGCCCGGGTCACCGTACGGGCCGTGGACGTCGCC
GACCGGGAGGCGCTCGCCGCCGCCCTGGCCACGGTGCCGGACGGCCACCCGCTCACCGGCGT
GGTGCACGCGGCGGGCATCCTGGACAACGGCCTGGTTTCCGCGCAGACGCCGGAGAGCCTGG
CCGCCGTGCTGCGGCCCAAGGCGGACGCGGCCTGGCATCTGCACGAGCTGACGAAGGACGTG
GACCTGTCGGCGTTCGTGCTGTTCTCGTCGAGCGTGGGCGTCGTCGGCGGGCCCGGCCAGTCC
AACTACGCGGCGGCCAACGCCTTCCTCGACGCGCTCGCCGAGCACCGGACCCGGCTTGGTCTT
CCCGCGAAGTCGCTCGCCTGGGGATTGTGGGACCTCGGCCACGGGATCAACGCGGGCCTGGA
CGCCAACGACCTCAAGCGGTTCACCCGGGAGGGGTTCCGTCAGGTCTCGCCCGAGGCGGGTA
CAGCACTGTTCGACGCCGCGCTGGACGGTTCCGCGGCGACGGTGGTGGCCCTGCCGGCCGAC
CTGTCCGCGATGCGGGCGCACGGCCGCGTCCCGGCGGTCTTCGGCGCTCTGGTGAGGGTGGC
GAACCGGCGCACCGCCCAGTCCGGGACGGTCGCGGCGGAGTCGTTCGCGCAGCGGCTTGCCG
GTCTGTCCGTACCGGAGCGGCGGCAGGCGGCCCTGGACCTGGTGCGGGTCGAAGTCGCCGCC
GCGCTGGGGCACTCGGGGCCGGACGCCGTGCCGGCCGAGCGTGCCTTCCAGGAGATGGGCTT
CGACTCGATGACCGCGGTCGAGCTGCGTAACCGGCTCATCGCCGCGAGCGGGGCACGGCTCG
CCGCCGCGGTGGTCTTCGACCACCCGACTCCGGAGGCCCTCGCCGAGCACCTTCTCGGCGAG
GTGGCACCCGAGGCGGCCGGCGACCCCGCGCAGCCGCTGCTGTCCGAACTGGACCGGCTGGA
GGCGTCGCTCTCGGCACTTGCCGGCGACGGCCGCGCCCGGTCCGCGGTCTCCGTCCGGCTCCA
GACGATCCTGTCCCGGCTGAACGAGGCCGCCGGACCGGACGAGAGTTCGGATGTGGTGAGCA
GCCTCGATTCCGCGTCGGCCGACGACCTCTTCGACTTCATCGACAACCAGCTCGGACGATCGG
CGAACTGA
5.SCO7221编码合成蛋白polyketide synthase,全长1170bp,具体核苷酸序列如下:
GTGGCCGCATACCTGTGCGCTCCCGCCGTGATACACGGCGAGCACTCCGTCGAGACCCGAGAG
ATCGTCGAGGAGGTACGCGGCCGGCACCCGCACGCTCCGTGGGCACCGCGGATCGACGGCAT
CGCGGCCAGTACGGGCATCGAGAGCCGCGGCTGGATGCTCCCCCTGGAGGCCGCCGTCGCGC
CCGGCGGTGGCGGCGACCTCGGGGCCGCGCGGGAGGCGCTGGTCCGCGACGGGTTCACCGAG
CAGGACGCGAACCGCGCGATCGCCGCCCTGAAGGCGGTGCCCGCGTCGCAGACCGTCCAGGA
GCGCACCGCCCCGGCCTGGGAGGCCGTGCAGGCCTACGGGGAGCGCGCGGCACGTGGTGCCC
TGCAGATCGCCGGGCTGGACGTCGCGGACGTCGACTGCCTGATCACCAGCAACTCCACCACCC
CGGCCCTGCCCGGACTGGACGTCGCCCTGGCCAACCGGCTCCCGCTGCGCGGCGACACCATG
CTCCTGCCCGCCACCCAGTGGGCCTGCGTGGCCGGGACCCGTTCCCTGGCACTGGCCGCCGAC
CTCGTGGCGGCGGACCCGGACCGGGTGGTCCTGGTCGTGATCTCCGAGGCGCTGAGCACCAC
CTACCAGCCCGCCGACGACACCCTGGAGTCCCTGATCGTCCGGCTGCTGTTCGCGGACACCGC
GGTCGCGGCGGTGGTCACGGGCCGCCCGAGGCCCGAGTCGGTGCTCCGGCTGGACGCCGCCT
GGCACCACACCCTGCCGGGCACGCGGGACCTGCACCGCCTGGAGACGCGGGCGGACGGCAC
CCACTTCGTGATGGACCGGCGCGGGCCGCGGGCGGTGCAGGAAACGGTCACCGCGATGTGGG
AGTGGCTCCGCGTCCGCTACGAGGACGACCCCTCCTCCTGGCACCCCGACGTCCTGCTCGCGC
ACCCCGGCGGGACCCGGGTGCTGGAGTACATGGAGCAGACGATGCCCGACGAGTGGCCCTCG
GGGCTCCTGTCGTACAGCCGGGACAGCTACACCAGCGGCAACCGCGGCGGCGCCGCCGTGTT
CGACATCCTGCGCCGGGCGCACGACGCCGGGCAGAAGACGGGCAGCCGTGCCGTCCTGTACG
CGGCGGCCCCGGGCCTGACCGCCACCGCTCTGGAAGGGGAGTGGCTGTAG
6.SCO7682编码合成蛋白non-ribosomal peptide synthase,全长6690bp,具体核苷酸序列如下:
GTGGAAGGAGACGCCGGCAACCGCGACCGCCACACCCCCGCCGCGGTACGGGAAGCCGTGG
CAGGCGCGCTGGAGATCGCACCGGAGGAGCTGGACGACGACCGCGACCTCTTCGAACTCGGC
CTCGACTCACTGGCGTTGATGTCCCTGGTCGGTACCTGGCGGCGCGAGGGCAGCACGGTCACC
TTCCAGGACCTCACGAGCAGGCCGACCCTGCGGGACTGGACGGCTCTGCTCGCGCGGGCCGC
GAACGACGACACGGCCACCACCGTTCCGCCCGGGCCACCCGCGCCGGTGACCGGTGCGGATC
CGGACACGGACTCCTTCCCGCTGGCCACGATGCAGCACGCCTACTGGATCGGACGCCAGGACG
GACAGCCCCTCGGCGGCGTCGCCGCGCACTTCTACGTCGAACTCGACGGCCACGGCGTCGAC
CCGGCCCGGCTGGACACCGCCCTGCGGTCGCTCGTGCTGCGGCACGGCATGCTCAGGGCCGTC
TTCGACGAGGACGGCCGTCAGCGCTTCGGGGCCGCCGGAGCGCCGCTCGCCGTCCACGACCT
GCGGGAACTCGCCCCCGTCGAGGCCGAGTCCGAGCTGGAGCGCCTGCGCGAGCGCAACACCC
ACACCCGCCCGGACATCGGCGCCGGTGACGTCTTCCGCACAGCCCTGTGCCTGCTCCCCGACG
GCAGGACCCGCCTCCAGATCAACCTGGACATGATGGCCGGAGACGCGCTGAGCCTGCGGGTC
CTCCTGTCCGACCTGCGCCGCTTCTACGAGTGCCCGGACGAACCGCCGCGCGAACTCGGGCTC
GACTACCGGCGCTACCTCGCGGAGCACGACGCCCGGCGACGGACCGAGCGAGAGCGGGACG
CCGCCTGGTGGCGCGAACGGCTGCCGGATCTGCCGCGGGCTCCGCGACTGCCGACGACCGTC
GACCCGCTGACCCCGGTCACCGCGGACGACGGCACGCTGACCCACTCCCGCCGCCTGCACCA
CTGGCTCGGCCCCGCCGACAAGGCCGCCCTCCTCGGCGCCGCCCGTCGGCACGCCGTCACCCC
CGCCGCCGCCCTGGCCACCGCCTTCGCCGAGGTGATCGCCGCCTGGAGCGACAGCCGGCGCTT
CCTCCTCAACCTGCCGCTCTTCGACCGGGAGATGTTCGCACCCGAAGTCGCCGGACTCGTCGG
CGACTTCAGCAGCTCGGTCCTGCTCGACGCGGACCTGAGCGCGCCCCTTGCCTTCTCCCGGCA
GGCCCGCGGCCTCCAGGAGCGGCTGCAGGCCGGAATCTCCCACGGGGCGTATGGCGGTGTCG
AGGTGCTGCGCGACCTCGCCCGCGCCGAGGGAGCACCCGTACTGGCCCCCGTCGTGTACACG
AGCGCCATCGGACTGGGGGAGATCTTCGAGCAGGACGTCCAGAAGTCCTTCGGCCACCCCTCC
TGGATCATCTCGCAGGGCCCGCAGGTGTATCTGGACGCCCAGGTCACCGAACTCGACGGCGGT
CTCCTCCTCAACTGGGACGTGCGGGACGGCGTGTTCGAGGCCGGCGTGCCCGACGCCGCGTT
CGACGCCTACCGCCGCCTCGTCACCCGCCTGGTGACCGACCCCGACGCCTGGGACCGGCCCGT
GGACGGCCTGCTCGCCCCCGGCGTCCTGCCCCGGCGCGCGGTGGCGGCCCTGCCCGCCGCGC
CCGTCGCGGACCGCGCCCTGCACACCCGCTTCTTCGGGCTGGCCGAGACCGAACCGGGCCGC
ACCGCGGTGGTCCGCACGACCGGCGACACCCTCACCTACGGCGAACTCGCCCACGCCGCACG
GCGGATCGCCGCGCTGCTGCACGACCACGGCGTGCGGGAGGGCGACACCGTCGCCGTCACCC
TGCCCAAGGGCGACCGGCAGATCACCGCGGTCCTCGGCATCCTGGCCGCGGGCGCGGCCTAC
GCCCCCGTCGGCGTGGAACAGCCCGCCGTGCGCCGGCAGCGCATCCACGCCGTGGCCGGCGC
CTCCGCCGTACTCACCGACCGGGACCACGCCCACCTGTGCGAGGCCGCCTCCCAGGCCCCGGT
GGTGCTCGTGGACGACGCCGCGGGACTCCGGCCCGCACCGGTCGTCCGGCCCGACCCCGGCC
TGGCCGCCTACGTGCTGTTCACATCCGGCTCGACGGGGCGGCCCAAGGCCGTCGAGGTGTCGC
ACCGGGCGGTCGTCAACACCGTCGAGGCGATGGAGGCACAGCTGGGGCTCGGCCCGGCGGAC
CGCACGCTCGCCATCTCCGCGCTGGACTTCGACCTCGCCACCTGGGACGTCTTCACCCCGCTG
TCCCTCGGCGGTCAGGTGGTGGCCGTAGGACAGGAACACCGCCGCGACGCCCACCACTGGGC
GCGGCTGGTGCGCACGCACGGGGTGACGCTGGTCCAGTGCGTGCCCGCCCTGCTGGACCTGC
TGATGGCCGCGGGGGAGGACGAGGGACTCGGGGACTCGCTGCGCATGGTGCTGCTCGGCGGC
GACTGGATCGGGCTCGACCAGCCCCGCAGACTCCGCGCGCTCGTCCCCGGCTGCCGTTTCGTG
GCGCTCGGCGGCATGACCGAGGCCGCCGTGCACTCCACGGTGTTCGAGGTGGAGGAGACCGA
CCCCGCCTGGAAGTCCGTCCCCTACGGCGTTCCGCTGCGCAACATGCGGGCCCGTGTCGTGGA
CGGACGCGGACGCGACTGCCCCGATCTCGTCCCGGGCGAGCTGTGGATCGGCGGCCCCGGCG
TGGCCAACGGCTACCGCGGGGACCCCGGACGCACGGCGGAACGCTTCGTCGACCACGACGGC
GAACGCTGGTACCGCAGCGGCGACCTGGCCCGCTACCGGCCCGACGGCGTGCTGGAGTTCCT
CGGCCGCGCCGACCACCAGGTGAAGATCGGCGGCCACCGCATCGAACTGGGGGAGGTCGAAT
CCGCGCTGGAGGACGACCCCGCCGTCCTGCACGCGGTCGCGACCGTCCTCGACACCCCCGTA
CGCCACCTCGCCGCGGCCGTGTCCGCCCCCGCCGGGGCACCGGCCCCGGACGAGATCCGCCT
GCGGGCGGCGGAACGCCTTCCCGCCCACATGGTCCCCGAGCGCGTCCTCGTCCTGCGCGACCT
GCCGCTGACCGCCAACGGCAAGCTCGACCGGACGGCCGTCCGGCAGGCGCTGACCGAGACCG
CCGGCCGGCCCGGACCCGCCACGTCGGCGCCGGCCGGCCCGGTGGAGACCGCCGTCGCGGCC
GAATGGGCCGACCTGCTCGGTGTGGCCGAAGTGGGACGCGAGAGCGGCTTCTTCCTGCTGGG
CGGCGACTCACTCGTCGCGACGCGCCTGATCGGCCGGCTGCGCGCCGCGGGCTTCCCGGGCG
CCCGCGTGGCCGACCTCTTCGCCCGCCCCGTACTCCGGCACTTCTGCGAGACGCTGCGCCCCG
CCACGACCACCGCCGAGGAGCCTGCCGCGGGCGCTCCTCCCGTCCTCGTGCCCGATCCGGACC
ACGCGCACGAGCCGTTCCCGCTGACCGACGTCCAGGCCGCCTACCACACCGGCCGCGACCCC
CGGTTCACCCTCGGCGGCGTGGGCACCTGGCACTACACGGAGTTCGACGGCACCGACGTCGA
CCTGGACCGGCTGGAGCGCGCCTGGAACACCCTGGTGCGACGGCACGGCATGCTCCGCGCCG
TCGTCGAGGACGGACACCAGCGTGTCCTGCCCGACGTACCGCCGGTGCGCATACCCGTGCACG
ACGCTCGAGCCGGTGGCGCGGCCGAGGCCCTGGCCGGCCTGCGCGCCCGCCTGTCCCAGCAG
GTCCGCGACCCCGCCCGCTGGCCGCTGTTCGCCATCGAGGCGGTGCGCCACCGCGCGGACGA
AGACACCGGCGAGAAGGGCGCCGAGACGCGGACCCGGATCGGCATCGGCCTGGACTACCTCG
TGCTCGACGCGCTGTCCATCACCACGCTCTACGCCGAACTGAACGCGCTCTACGCCGACCCGG
AACACCAACTTCCCCCCGTCGGCGTGTCGTTCCGCGACTACCTGACCCGCCTCCCGGCCGACC
CCGAGTCGACCGTGCGGGCCCGCGCCCACTGGCAGGCCCGCCTGCCGGAACTGCCACCGCCG
CCCGCCCTCCCCTTCGACCGGGACCCGGCGACGCTCGACGCCCCCGTCTTCACCCGGCGCCGG
CTGAGCCTGCCGGCCGGGGACTGGCAGGCCGTCAAGCGGGACGCCGCCCGGCACGGCCTGAC
CCCGTCCACCGTCCTGCTCGCCCTGTACGCGGAGGTCCTGGCGGCCTGGAGCGGGGCCGACGC
CGTGACCGTCACGCTCACCCTCTTCAACCGGCGCGACGTCCACCCCCACATCCACCGGGTACT
GGGCGACTTCACCTCCCTGTCCCTCGTCGGCTACCGGCGCGAGGGCGCCGACTGGCCCGCGGC
GGCCGCCGAGCTGCAACGCCGCCAGGCCGAGGACCTCGACCACGCCGACGTGCCGATCGACT
GGCTGCTGCGGGAGTTCGGCCGGCGCACGGGCACGGTGGACGCCGCCGCACCCGTGGTGTTC
ACCAGCGCCATCGGCGTCGGGGACGCGACGCTGGCCGGACCGGGCACCGGACTCCCCCCGAA
GGTCTGGGGCATCTCCCAGTCGCCGCAGGTCTGCCTGGACAACCAGGTCACCGAGGAATCCG
GGAACCTCGTCGTCACCTGGGACGCCGTCGAGGAACCGTTCGCCGAGGGTGTGCTGGACGCG
ATGTTCTCCGCGTACGAACGGCTGCTGCGGCACCTCGTGGACGGTGACTGGCAGGCACCCCTC
CCCGACCCGGTGCCCGCCGCCCAGTACGAACGGCGGGAGGCCGCGGCACGGGAGGCCGCCCT
GGAACCGGTGACACCGCGCCCGCTGCACCAGACGTTCTTCACCCACGCGGCGGCCACCCCCG
ACCGCACCGCCCTCGTCACGACGGCGGGGGAGGAGACGACCTACGGAACCCTCGCCGCACAG
GCGCTGCGCACCGCCGCCGCGCTCGCCGCCGCCGGGGTGCGGCCCGGCGACCTGGTCGCGGT
CACCCTGCCCAAGGGCCCGGAACAGGTCGCGGCGGTCCTCGGCACGCTCGCGGCCGGCGCCG
CCTACGTCCCCCTGAGCCTCCAACAGCCCGCCGCACGGCTGGAGCGGGTGCGCGCCACGGCC
GGTTTCGACGTCGTCCTGGGCGGCTGGCCCGCCACGGACGGCGCCGGCCCCCGCACCCTGTCC
CCGGCCGACACGCAGGGCGCCGAGCCGCTGTCCACGCCGGTCGGAACGGCTCCCGACGCGCT
CGCCTACGTGATCTTCACGTCGGGCTCCACCGGCGAGCCCAAGGGCGTCGAGATCACCCACGC
CGCCGCCTGGAACACGATCGCGGACATCAACGCCCGCCACGCCATCCGGGGCGACGACCGCG
TCTTCGCCCTGTCGGCACTGGACTTCGACCTGTCCGTGTACGACGTCTTCGGCCTGCTCGCCGC
GGGCGGCTCCCTCCTGCTGCCCACCGAGGACCAACGCCGTGAACCGGCCCGCTGGCCGGACC
TGATGGACCGGTACGGCGTGACCGTCTGGAACACCGTCCCCGCCCTGCTCGACCTGCTGCTCG
ACGCCGACGGACAGGAGGGACGGGAGGGCCGCCGGATCGCGGGGCTGCGAACCGCCCTGGT
CTCCGGCGACTGGATCGGCCTGGACCTGCCCGCCCGGCTGCGCGCCCGAACCGGGCGGTCCT
GCGCCTTCGTCGCCATGGGCGGAGCCACGGAAGCCGCCATCTGGTCCAACACCCTCACCGTCA
CCGAACCCGACCCCCGGTGGGTGTCCATCCCCTACGGCCGCCCCCTGACCGGCCAGCACTACC
GCGTGGTCGACCGGGACGGCCGCGACTGCCCCGACTGGACGCCCGGCGAGCTGTGGATCGGC
GGCGCGGGACTGGCCCGCGGCTACCTGGCCGACCCGGCCCGCACCGCCGAGAAGTTCGTCGA
ACGGGACGGGCGGCGCTGGTACCGGACAGGAGACCTGGGCCGGTTCCGGGACGACGGACTG
CTGGAGTTCCTCGGCCGGCTCGACAGCCAGCTGAAGATCGCCGGCCACCGCATCGAGGCCGG
CGAGGTCGAGGCCGCGCTGGAGGCCCACCCGGCCGTCGCGCGCGCCGCCGCCCTGGCCGTGG
GGGAGCGCACCGCCAGACGCCTGGTGGCGTTCGTCGTACCGCACGAACCACCGCCCGCCGAC
GAAGCCGAGCCGGTCGAGAACCCCGGACTCACCGAGGACCTCCTCGGCCTGCTGGCCGAGCG
GGTACCGGTCTACGCCGTCCCGTCCCGCATCCTGTCCCTGCCCGCACTGCCCCTCACCGCGAAC
GGCAAGGTCGACCGGGCCGCCCTGACCGGCCTCGCGGGACCCGAGGCGGTCTCCACCGCCGC
CGAGCCGCCACGCGGCGAGACCGAGACGGCCCTCGCCGCCCTCTGGCACGCATGCCTGCCGC
ACGGGTCGCCGGACCGCAACGCCAACTTCTTCACCGCCGGCGGCGACAGCCTCACCGCGCTG
CGCCTGCTCACGGCGATCGAGCGCCGCTTCGGGGCCGCCGTCCCCGCCCGCCGCTTCTTCGCC
GCACCGACCATCGCCGCGCTCGCCGCGGACATCACCCGAACGCTCGCGGCCCACGGGACCGC
CTACGAGACCGGAGAGCTGTGA
7.SCO7683编码合成蛋白non-ribosomal peptide synthase,全长5529bp,具体核苷酸序列如下:
GTGAACATCGCTGAACTGCTGGCCCGATACGCGGACGAAGGCGTGGTGCTGTGGGCCGA
GGACGGACAACTGCGGTTCCGGGCCCCGCAGGGCCGGCTCACCGACACGCACCGAGCCGAAC
TGCGCACGCACAAGGAAGCCGTCCTGCGCCATCTGGAAGCGGCGGAACCGGCGCTGCGGGCC
GACCCCGACCACCGCCACGAACCCTTCCCCCTGACGGACATCCAGGCCGCCTACCTCATCGGC
CGCACCGACGCCTACGCCTACGGGGGCGTCGGCTGCCACGCCTACGTGGAGCTGGCATATCCC
GACCTCGACGTCGACCGAGTCACCTCCGTGTGGCGGCAACTGGTCCAGCGGCACGACATGCT
CCGGGCCGTCATCCACCAGGACGGCCACCAGCGGGTCCTGCCCGAGGTGCCCGCGCTGTCCG
TCGGGACCGACGACCTGACCGCCCTGCCGGCCGCCACCGCCGAGGCACGGATGCGGCACACC
CGGGCGCGGCTGTCCGCCCGCGAGGCGCCCACCGACCAGTGGCCCCTCTTCGACGTACACGT
GACCCGGACCGACCGGCGGGCCGTACTGCACCTCTCCTTCGACATGCTCGTCGTCGACCACGC
CAGCCTGCGGATCCTGCTCGCCGAGTTCCGGCGGAGCTACGGCGGCACCGCCCTCACCGACGC
GCCGCGGATCACTTTCCGCGACTACGTCCTGGCCCGCCGCGCCCTTGCCGACACCGACGGCCA
CGCCCGCGACCGCGCCTACTGGACCGAACGGCTCGACACCCTGCCGCCCGCCCCCGAACTGC
CCCTCGCCGAAGCCTGGCAGGCCGCGGTGGACGACCCGGCGGAAGCGGGCCGCGTCCCGGCG
CCCGGCGGCGACGCGGTGGCGTTCCGCCGGCTGGAGGTGCTGCTGCCCGCGGCCGACCGGGA
CCGGCTCACCGCACGTGCCGCGCGGCGCGGGCTGACCCCGTCGACGGCCCTGCTCACCGCTTA
CGCCGAGACGGTCGGCGCCTGGTCGCGCTCCAGCCGGTTCACCCTCAACGTGCCCACCGTCG
ACCGGCCCGCCCTGCACGAGGACATCGACCGCCTCGTCGGCGACTTCACCTCGCTCGAACTCC
TCGCCGTCGACCTCGACACCCCGGCCACCTTCGCCGAACGGACCGGGGCCGTGGGCGAACAG
CTCCTCGATGACCTGGCCCACCCGCTGTTCACCGGCTCGGAGGTGCTCGCCGAACTCTCCCGC
AGGGCCGGCGCTCCGGTCCTCATGCCGGTCGTCTTCACCAGCGCCCTGGGCGCGGGTGCCACC
TCCGAGGGCGTACCCCCCGAGGTGGAGTACGCCGTGACCCGGACACCGCAGGTGTGGCTCGA
CTGCCAGGTCATGCACCGCGGCGACACCCTCAGCCTGTCCTGGGACATCCGGGAGGGCGCGCT
CGCCCACGGGACGGCCGACGCCATGTTCGAGGCGTACACGGCACTCGTGCGTTCGCTGTCGGC
CGAGGGCGAGGCAGGCGAGAAGGCCTGGGACGCACCCGTCCGCATCCCGCTGCCCGCGGCAC
AGGCGGCACGCCGGGCCGCGGTCAACGCGACCGAGGGCCCCCTCCCGGACGCCCTGCTCCAC
GAACCCGTACTCGCCCGGGCACGGACCACACCGGACGCGATCGCGGTCCGCACCCCCGAACT
GGCCCTCTCCTACCGGCAGCTCGTCACCCGCGCCACAGGACTCGCCCAGCACCTGACGGCCTC
CGGACTGCGCCCCGGGGAACCGGTGGCCATCTGGATGGACAAGGGCTGGGAACAGGTCGTCG
CCGTCTTCGGCACCCTCATGGCGGGCGGCGCCTACCTGCCCGTCGACACGGCCCAGCCGGCGG
CCCGCCGGGACACGATCATCGGGGACGCGGGCGTCCGCACCGTCCTCACCCAGTCCTGGCTCG
CCGAACTCGAAGACCTCCCCTCGACGGTGTCGCCGGTCGCCGTGGACCTCGTCGGGGAGGCC
ACCGCGGACCTCCCGCCCGCCGCCCGCCGGGACCCTGACGACCTCGCCTACGTCATCTACACC
TCGGGCTCCACCGGCACGCCCAAAGGCGTGATGATCAGCCACCGGGCCGCCCTCAACACCGT
CGAGGACATCAACCGCCGCTTCGCCGTCGACGAGCGCGACCGCGTCCTCGGCATCGCCGGACT
CGGCTTCGACCTGTCCGTCTACGACCTGTTCGGACCGCTCGCCGTGGGCGCCACCCTCGTCCT
GCCCCGGTCCGACCTGCGCGGCGACCCCTCCCACTGGGCCGAACTCGTCCGCGACTTCGGCGT
CACGGTGTGGAACTCCGTACCGGGCCAGCTCCACATGCTGTGCGACTGGCTCCGCTCCGAGCC
CCCCACCGACGACGGCTCCCTGCGCCTGGCACTGATATCGGGCGACTGGATACCGGTCGCACT
GCCCGACCAGGCCCGTGAACTCCTGCCCGGACTGGAGATCGTCTCCCTCGGCGGAGCCACCG
AGGGCTCCATCTGGTCCATCGCCCACCCGATCGGCGAGGTCGACACCGCCCGCCCCAGCATCC
CGTACGGCAAGCCCCTGACCAACCAGACGTTCGCCGTGCTCGACCGGCACCTGCGGCCGCGG
CCCGAGTGGGTGCCGGGCGAGCTGTACATCGGCGGTGCCGGTGTCGCCCTGGGCTACCTCGGC
GACGGCGAGCGCACGGCACAACGGTTCCTCACCGACTTGGCCACCGGAGAGCGGCTCTACCG
CACGGGCGACCTCGGCCGCTACCTGCCCGACGGGACCATCGAGTTCCTGGGCCGGGAGGACG
CCCAGATCAAGATCCGCGGATACCGGGTCGAACTCGCCGAGGTGGAGGCCGCCGTACAGACC
CACCCCGCGGTCGCCGCGGGCGCGGTCGTCGTGGACGACTCGGCGGCGGGCGGGCGGCGCCT
GGCCGCGTTCGTCGAGACGGCACGCAAGGACGGCGGGCCCGCCGTGACGAGCCGCGCACAG
GCGCGCACGGCAGCCGCCGAAGCGGTACGCGAAGCGGGCGCGGCCGTCGACGCCGCGCGGC
TCACCGACTTCCTCGCCGCCCTCGACGACGTTGCCGTCGCCGAGATGACCCGGGTCCTCGCCG
CCTCCGGAGTGTTCGACTCGGCCGCGGCACGCACCGCGGAGGAGGTGGGCACGGCACTGCGC
GCCACACCGCGCCACCGGCACATCGTGCGCCGCTGGCTGCGCGCCCTCGCCGCCCGGGACCG
GCTCACCCACCGGGACGCCGACGCGACCTACACCGGCCTGCGCCCCGTCTCCGCGCCCGAGG
CCGAACGCCGGTGGCGGCGGGCCGCCGACCTGGAGCACGAGATCGGGTGGAGCACCGAACT
GCTCACCGTCATGCGCACCTGCGCCGAGCGGCTGCCCGAGCTGGTCTGTGGGGACGTCGGCAT
CCGGGACCTGCTCTTCCCGGGCGCGGCGACGGAGGCCGCGGACGCCGCCTACCGGGACAACC
TCGCCATCCGGCACCTCAACCGCGCCGTGGTCGCCGCGCTGCGCGAGATCGCCGCGGGCCACA
CCGGCGAGGAGCGGCTGCGGGTGCTGGAGGTCGGCGGCGGCGTCGGCGGTACCACCGGCGA
ACTGGTGCCCGTCCTCGCCGAGTACGGCGTCGACTACCTGTTCACCGATCCCTCGGCGTTCTTC
CTGAACGAGGCCCGCGAGCGCTTCGCCGACCACGCCTGGGTCCGCTACCAGCGCTTCGACGTC
GACGAGGACCCGCGCGCCCAGCACCTGCTGCCCAACACCTTCGACGTCGTGGTCTGCGCCAA
CACGCTGCACGCGGCAGCCGACGCGGACGCCGCCGTGGGCAGGCTGCGGGAACTGCTCGTAC
CGGGCGGACAGCTCGTGTTCGTCGAGAACACCCGCGACGAGAACCTCCCGCTGCTGGTCTCC
ATGGAGTTCCTGGAGGTCGCCGGCCGGACCTGGACCGACGTGCGCGAGCACACCGGGCAGTC
CTTCCTCACCCACACCCAGTGGCGTGCCCTCCTCGACGGGCACGAGGCGTCCGGGGTCACCTC
GCTGCCCGACGCCGGGGACGCCCTGGCCGGCACCGGACAGGAGGTGTTCATCGCCACGATGA
AGGACGACCGGCACCACGTGCCGGTGGGGGAACTGGCCCGAACGGCGGCCACCCGGCTCCC
GGAGTACATGCTCCCCGCGGTGTGGCAGGTGGTCGACGCGCTGCCCCGCACCGCGAACGGCA
AGACCGACCGCGCGCGCCTGCGGTCCTGGCTGCCCCGGGAGAGCGCACCGGTCGTCGTGGAC
GAGCAGCCGAAGGACGAGCTGGAGCACAGCCTGGCCGACCTGTGGGCGGAACTCCTCGCGGT
CGAAGGCGTCACCCGCAACGACGACTTCTTCGACCTGGGCGGCGACTCGCTCCTGGTGGCCC
GGATGGTGGGCCGGCTGCGGGAACGCGTACCGCAGGCGGCGGACCTGGAGTGGGAGGTCGTC
CTGCGCCACATGCTGCGCCGCCCCACGGTCGCGGGTCTCGCCGCGTTCCTGCGCGCTCTGGCC
GGTCCCGAGGACGCGCCCGCGGCCGGTGCCGTACGCACGGACCCGGTGATCCACCTGCACGG
CTCCCGCTCCGAGGACGAGCCGACGACCGTGCTGGTGCACGCCGGAACCGCCACGATCATGC
CGTACCGGGCGCTGATCACCGAGATCAGGCGCCGCTCCCCGGGCCTCGCCGAGGTCGTCGGTG
TGGAGGTCCCGGACCTGAGCGGATTCCTCGCCGCACCCCCGGACGGACTCATCGAGCGGATGG
CGGCCGACTACGCACGGGCGCTGACGGCGGACGGCCGCCACCGCTTCCACGTCGTCGGCTAC
TGCCTGGGCGGCCTGATCGCCACCGAAGTGGCCCGCAACCTGGCCGAGTCGGGCGCGGAGGT
GGAGAGCCTCACCGTCATCAGCAGCCACAGCCCCCGCTTCCGCCTGGACGACGAACTCCTCGC
CGAGTACTCCTTCGCCGTCATGATGGGCATCGACCCGGCCGACCTCGGGTTCCCGGACGACCA
GTACCGGGTGGCCGCCGCGGCCGACACCGTCCTGGCCGCCAGTCCCGGCGTCCTGCCGGACG
GCGGACTCGCCGCACTGGGCGGGGAGTTCGAGGACGTCGCGAGCCGCTTCCGGCGACTCGCC
GACGTCCCCCGCGCCACCCGTATCGCCCGGATGTGCGAGGCGGTTCCCGCGTCGGCCGGCACC
TACGAACCCGACCACATGACACGGCTCTTCCTCGCCTTCCGCCAGAGCGTCTTCGCCATCACC
CGCTACCGGGCGGAGCCCTACGCGGGTGACATCACCTTCCTGCGCCACGACGGCGCCTACCCC
TTCCCCGGCAGCAAGGACGCCGTCACCGCCTACTGGGAGGAACTGACCCTGGGAGACCTGGA
CATCGTCGACATCGGCGGAGACCACTACAGCTGCCTGTCCGTCGAACACGCGCCCGGGATCCT
CAAGACCCTCGGCGAGCTGACCCAGGGGGCGATCACCCGATGA
在本发明的方案中,本发明发现了不同来源的细菌、真菌信号分子可以激活天蓝色链霉菌A3(2)关键次生代谢产物的表达上调。
与现有技术相比,本方案的有益效果:
本发明的方法通过常见的代表性群体感应信号分子Farnesol、Tyrosol、3-oxo-C12-HSL和Tryptophol对天蓝色链霉菌A3(2)(模式放线菌研究模型)沉默BGCs激活的作用,开发了一种新型信号分子激活放线菌沉默基因簇的表达的方法,即为一种快速、高效激活不同放线菌中沉默BGCs的策略,本发明的该方法将为充分发掘新型放线菌天然产物提供有效途径,将为临床药物研发贡献更多更好的新型先导化合物。
附图说明
图1是本发明实施例中天蓝色链霉菌A3(2)基因组信息中次生代谢产物合成基因簇预测结果示意图;
图2是本发明实施例中正常条件下天蓝色链霉菌A3(2)的生长曲线;
图3是本发明实施例中通过白色念珠菌信号分子Farnesol和Tyrosol激活天蓝色链霉菌A3(2)沉默基因簇表达的qRT-PCR结果示意图;
图4是本发明实施例中通过铜假单胞菌信号分子3-oxo-C12-HSL激活天蓝色链霉菌A3(2)沉默基因簇表达的qRT-PCR结果示意图;
图5是本发明实施例中通过酿酒酵母信号分子Tryptophol激活天蓝色链霉菌A3(2)沉默基因簇表达的qRT-PCR结果示意图。
具体实施方式
为了使本技术领域的人员更好地理解本发明方案,下面将结合本发明的实施例及附图,对本发明的技术方案进行进一步详细地描述,显然,所描述的实施例仅仅是本发明一部分的实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都应当属于本发明保护的范围。
需要说明的是,在不冲突的情况下,本发明中的实施例及实施例中的特征可以相互组合。下面将结合实施例来详细说明本发明。
实施例1:天蓝色链霉菌A3(2)基因组信息中次生代谢产物预测
将天蓝色链霉菌A3(2)基因组信息GCA_000203835.1_ASM20383v1利用antiSMASH在线预测网站预测分析。发现至少有18种次生代谢产物,其中至少有12种可能属于PKS或NRPS类抗生素。
实施例2:天蓝色链霉菌A3(2)生长测定
培养至稳定期的天蓝色链霉菌A3(2)按1:100转接至发酵培养基中,定点取样,采用称取链霉菌细胞干重的方法,测量并绘制链霉菌的生长曲线图。其中,48h后链霉菌开始进入生长对数期,在96h左右到达对数末期,120h后进入稳定期。
实施例3:天蓝色链霉菌A3(2)中次生代谢基因簇的功能基因的定量分析
将天蓝色链霉菌A3(2)在发酵培养基中30℃培养至对数中后期,分别加入细菌、真菌相关信号分子(100μM Farnesol、100μM Tyrosol、100μM 3-oxo-C12-HSL、100μMTryptophol)培养2.5h后,离心收集菌体。
(1)天蓝色链霉菌A3(2)总RNA提取,使用RNA提取试剂盒(天根生物科技,中国),具体操作如下:
①用含有10mg/mL溶菌酶的100μL dd H2O彻底重悬菌体,室温孵育15min;
②加入350μL裂解液RL(在使用之间已加入β-巯基乙醇),涡旋振荡混匀;
③加入250μL无水乙醇。将得到的溶液和沉淀一起转入吸附柱CR3中,12000rpm离心1min,倒掉废液,将吸附柱放回收集管中;
④向吸附柱CR3中加入350μL去蛋白液RW1,12000rpm离心1min,弃废液,将吸附柱放回收集管中;
⑤DNase I工作液的配制:取10μL DNase I储存液放入新的RNase-Free离心管中,加入70μL RDD缓冲液,轻柔混匀;
⑥向吸附柱CR3中央加入80μL的DNase I工作液,室温放置15min;
⑦向吸附柱CR3中加入350μL去漂洗液RW1,12000rpm离心1min,弃废液,将吸附柱放回收集管中;
⑧向吸附柱CR3中加入500μL去蛋白液RW(使用前已加乙醇),室温放置2min,12000rpm离心1min,弃废液,将吸附柱放回收集管中,并重复一次;
⑨12000rpm离心2min,倒掉废液,将吸附柱CR3置于室温放置10min;
⑩将吸附柱CR3转入一个新的RNase-Free离心管中,向吸附膜的中间部位悬空滴加50μL RNase-Free dd H2O,室温放置2min,12000rpm离心2min,得到的RNA样品利用琼脂糖凝胶电泳检测RNA的完整性和纯度,采用酶标仪测定浓度。
(2)RNA反转录和定量分析
首先按照TransGen公司反转录试剂盒进行操作。按照表1反转录体系和表2反转录程序产生cDNA,然后按照表3的定量体系、表4的定量相关引物和表5的定量程序,利用LightCycler 96热循环仪(Roche,美国)进行qRT-PCR反应。其中,使用天蓝色链霉菌A3(2)的hrdB基因作为内参。
表1RNA反转录体系
表2RNA反转录程序
表3定量体系
表4定量相关引物
表5定量程序
通过本发明的上述实施例,本发明通过研究群体感应信号分子Farnesol、Tyrosol、3-oxo-C12-HSL和Tryptophol对天蓝色链霉菌A3(2)(模式放线菌研究模型)沉默BGCs激活的作用,开发了一种新型信号分子激活放线菌沉默基因簇的表达的方法,其为一种快速、高效激活不同放线菌中沉默BGCs的策略,将为激活大量存在的沉默BGCs宝库,深入发掘新型放线菌天然产物提供有效途径,从而为医药、农业等多领域的研发贡献更多更好的新型先导化合物。
以上具体实施例仅仅是对本发明的解释,其并不是对本发明的限制,本领域技术人员在阅读完本说明书后可以根据需要对本实施例做出没有创造性贡献的修改,但只要在本发明的权利要求范围内都受到专利法的保护。
Claims (5)
1.一种激活放线菌沉默次生代谢产物合成基因簇表达的方法,其特征是:所述方法为:采用代表性微生物群体感应信号分子刺激放线菌沉默基因簇,使放线菌沉默BGCs被激活;其中,所述的微生物群体感应信号分子以Farnesol、Tyrosol、3-oxo-C12-HSL和Tryptophol作为代表但并不局限于以上四种。
2.如权利要求1所述的一种激活放线菌沉默次生代谢产物合成基因簇表达的方法,其特征是:采用代表性微生物群体感应信号分子刺激供试放线菌菌株天蓝色链霉菌A3(2)。
3.如权利要求1-2任意所述的一种激活放线菌沉默次生代谢产物合成基因簇表达的方法的应用,其特征是:将所述方法应用于供试放线菌代表菌株天蓝色链霉菌A3(2)次生代谢产物预测分析,并应用于潜在的抗生素合成。
4.如权利要求1-2任意一项所述的一种激活放线菌沉默次生代谢产物合成基因簇表达的方法的应用,其特征是:将所述方法用于获得新型放线菌次生代谢产物编码合成蛋白的核苷酸和抗生素的生物合成。
5.如权利要求4所述的应用,其特征是:编码所述合成蛋白的核苷酸序列如SEQ ID NO:2所示。
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