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菌物学报  2015 

高通量测序检测金针菇RNAi转化子插入位点及拷贝数

DOI: 10.13346/j.mycosystema.150089, PP. 694-702

Keywords: 基因组重测序,农杆菌介导转化,金针菇,高通量测序

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Abstract:

本研究对金针菇Flammulinavelutipes的一个RNAi转化子菌株1382R3进行了高通量测序,以本实验室先前获得的野生型W23基因组数据为参考,分析了该转化子的基因插入位点以及拷贝数。转化子菌株1382R3是通过农杆菌介导将fv-hmg1-RNAi载体转化至金针菇菌株并通过PCR检测筛选标记而得到。通过BLAST将转化子测序的reads对外源载体和基因组定位,找到具有基因组序列(GS)和外源载体序列(ES)两种序列的临界reads,并据此使用PERL语言程序成功在转化子1382R3菌株中找到两个插入位点。对两个插入位置的序列分析表明:在插入位点1,T-DNA片段部分插入;在插入位点2,T-DNA全部插入到基因组。两个插入位点都对基因组内源基因的表达造成了一定的干扰。此方法拓宽了高通量测序技术的应用范围,将其运用到遗传转化插入位置和拷贝数的研究中,有利于食用菌的功能基因组及基因工程研究。

References

[1]  Zhang F, Jin WZ, Chen SY, Wu YR, Liu FY, Wu P, 2004. Comparison of the efficiency between plasmid rescue and TAIL-PCR methods for obtaining flanking sequence of T-DNA tag insertion in rice genome. Journal of Agricultural Biotechnology, 12(1): 13-18 (in Chinese) [本文引用:1]
[2]  陈松, 张洁夫, 浦惠明, 申爱娟, 周小樱, 龙卫华, 胡茂龙, 戚存扣, 2011. 转基因高油酸油菜T-DNA插入拷贝数及整合位点分析. 分子植物育种, 9(1): 17-24 [本文引用:2]
[3]  樊荣辉, 武治印, 王永强, 赫荣琳, 林福呈, 王洪凯, 2010. RNAi技术及其在真菌基因功能研究中的应用. 菌物学报, 29(3): 315-320 [本文引用:2]
[4]  李彩红, 李志芳, 冯自力, 赵丽红, 师勇强, 王玲飞, 刘义杰, 朱荷琴, 2013. 农杆菌介导的基因敲除技术在丝状真菌基因功能研究中的应用. 棉花学报, 25(3): 262-268 [本文引用:1]
[5]  李今煜, 陈健铭, 彭振坤, 2007. 几种常用的基因敲除技术. 武夷科学, 23(1): 187-190 [本文引用:1]
[6]  梁成真, 张锐, 郭三堆, 2009. 染色体步移技术研究进展. 生物技术通报, 30(10): 75-83 [本文引用:1]
[7]  刘芳, 王威, 谢宝贵, 2014. 金针菇菌丝与原基差异表达基因分析. 食用菌学报, 21(1): 1-7 [本文引用:1]
[8]  罗滨, 陈永康, 王莹, 2012. 植物外源基因拷贝数及插入位点的检测方法与技术. 河南师范大学学报(自然科学版), 40(6): 111-116 [本文引用:1]
[9]  施乐乐, van Peer AF, 郭丽, 陈仁良, 王威, 严俊杰, 邓优锦, 谢宝贵, 2014. 农杆菌介导一个内源HMG-box转录因子fvhom1转化金针菇的研究. 基因组学与应用生物学, 3(6): 1268-1274 [本文引用:4]
[10]  王凯, 张燕洁, 关兵, 郭旺珍, 张天真, 2007. 棉花细菌人工染色体的荧光原位杂交(BAC-FISH)技术. 生物化学与生物物理进展, 34(11): 1216-1222 [本文引用:1]
[11]  王兴春, 杨致荣, 王敏, 李玮, 李生才, 2012. 高通量测序技术及其应用. 中国生物工程杂志, 32(1): 109-114 [本文引用:3]
[12]  翟志芳, 许文涛, 张南, 闫兴华, 王一南, 罗云波, 黄昆仑, 2011. 转基因玉米LY038转化事件的特异性检测. 农业生物技术学报, 9(3): 577-582 [本文引用:1]
[13]  张帆, 金维正, 陈双燕, 吴运荣, 刘非燕, 吴平, 2004. 质粒营救法和TAIL-PCR法获得水稻T-DNA旁邻序列的效率比较. 农业生物技术学报, 12(1): 13-18 [本文引用:1]
[14]  Wang XC, Yang ZR, Wang M, Li W, Li SC, 2012. High-throughput sequencing technology and its application. China Biotechnology, 32(1): 109-114 (in Chinese) [本文引用:1]
[15]  Wang W, Liu F, Jiang Y, Wu G, Guo L, Chen R, Chen B, Lu Y, Dai Y, Xie B, 2015. The multigene family of fungal laccases and their expression in the white rot basidiomycete Flammulina velutipes. Gene, 563(2): 142-149 [本文引用:1]
[16]  Zhai ZF, Xu WT, Zhang N, Yan XH, Wang YN, Luo YB, Huang KL, 2011. Event-specific transgenic detection of genetically modified maize LY038. Journal of Agricultural Biotechnology, 9(3): 577-582 (in Chinese) [本文引用:1]
[17]  Altschul SF, Madden TL, Sch?ffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ, 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research, 25: 3389-3402 [本文引用:1]
[18]  Bao D, Gong M, Zheng H, Chen M, Zhang L, Wang H, Jiang J, Wu L, Zhu Y, Zhu G, Zhou Y, Li C, Wang S, Zhao Y, Zhao G, Tan Q, 2013. Sequencing and comparative analysis of the straw mushroom (Volvariella volvacea) genome. PLoS One, 8(3): e58294 [本文引用:1]
[19]  Chen B, Gui F, Xie B, Deng Y, Sun X, Lin M, Tao Y, Li S, 2013. Composition and expression of genes encoding carbohydrate-active enzymes in the straw-degrading mushroom Volvariella volvacea. PLoS One, 8(3): e58780 [本文引用:1]
[20]  Chen S, Xu J, Liu C, Zhu Y, Nelson DR, Zhou S, Li C, Wang L, Guo X, Sun Y, Luo H, Li Y, Song J, Henrissat B, Levasseur A, Qian J, Li J, Luo X, Shi L, He L, Xiang L, Xu X, Niu Y, Li Q, Han MV, Yan H, Zhang J, Chen H, Lü A, Wang Z, Liu M, Schwartz DC, Sun C, 2012. Genome sequence of the model medicinal mushroom Ganoderma lucidum. Nature Communications, 3: 913 [本文引用:1]
[21]  Chen S, Zhang JF, Pu HM, Shen AJ, Zhou XY, Long WH, Hu ML, Qi CK, 2011. Analysis of transgene copy and integration site of T-DNA to genome of transgenic high oelic (Brassica napus L. ). Molecular Plant Breeding, 9(1): 17-24 (in Chinese) [本文引用:1]
[22]  Cho JH, Lee SE, Chang WB, Cha JS, 2006. Agrobacterium-mediated transformation of the winter mushroom, Flammulina velutipes. Mycobiology, 34(2): 104-107 [本文引用:1]
[23]  De Preter K, Speleman F, Combaret V, Lunec J, Laureys G, Eussen BHJ, Francotte N, Board J, Pearson ADJ, De Paepe A, Van Roy N, Vand esompele J, 2002. Quantification of MYCN, DDX1, and NAG gene copy number in neuroblastoma using a real-time quantitative PCR assay. Modern Pathology, 15(2): 159-166 [本文引用:1]
[24]  DiCarlo JE, Norville JE, Mali P, Rios X, Aach J, Church GM, 2013. Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic Acids Research, 2013: gkt135 [本文引用:1]
[25]  Fan RH, Wu ZY, Wang YQ, He RL, Lin FC, Wang HK, 2010. RNAi technology and its application to gene function of fungi. Mycosystema, 29(3): 315-320 (in Chinese) [本文引用:1]
[26]  Fitzgerald AM, Mudge AM, Gleave AP, Plummer KM, 2003. Agrobacterium and PEG-mediated transformation of the phytopathogen Venturia inaequalis. Mycological Research, 107(7): 803-810 [本文引用:1]
[27]  Gabriel R, Eckenberg R, Paruzynski A, Bartholomae CC, Nowrouzi A, Arens A, Howe SJ, Recchia A, Cattoglio C, Wang W, Faber K, Schwarwaelder K, Kirsten R, Deichmann A, Ball CR, Balaggan KS, Yá?ez-Mu?oz RJ, Ali RR, Gaspar HB, Biasco L, Aiuti A, Cesana D, Montini E, Naldini L, Cohen-Haguenauer O, Mavilio F, Thrasher AJ, Glimm H, von Kalle C, Saurin W, Schmidt M, 2009. Comprehensive genomic access to vector integration in clinical gene therapy. Nature Medicine, 15(12): 1431-1436 [本文引用:1]
[28]  Gase K, Weinhold A, Bozorov T, Schuck S, Baldwin IT, 2011. Efficient screening of transgenic plant lines for ecological research. Molecular Ecology Resources, 11(5): 890-902 [本文引用:2]
[29]  Inglis PW, Aragao FJL, Frazao H, Magalh?es BP, Valadares-Inglis MC, 2000. Biolistic co-transformation of Metarhizium anisopliae var. acridum strain CG423 with green fluorescent protein and resistance to glufosinate ammonium. FEMS Microbiology Letters, 191(2): 249-254 [本文引用:1]
[30]  Kohda T, Taira K, 2000. A simple and efficient method to determine the terminal sequences of restriction fragments containing known sequences. DNA Research, 7(2): 151-155 [本文引用:1]
[31]  Kuo CY, Chou SY, Hseu RS, Huang CT, 2010. Heterologous expression of EGFP in enoki mushroom Flammulina velutipes. Botanical Studies, 51(3): 303-309 [本文引用:2]
[32]  Kuspa A, Loomis WF, 1992. Tagging developmental genes in dictyostelium by restriction enzyme-mediated integration of plasmid DNA. Proceedings of the National Academy of Sciences, 89(18): 8803-8807 [本文引用:1]
[33]  Li CH, Li ZF, Feng ZL, Zhao LH, Shi YQ, Wang LF, Liu YJ, Zhu HQ, 2013. Appliacations of gene knockout by ATMT in filamentous fungus functional genomics. Cotton Science, 25(3): 262-268 (in Chinese) [本文引用:1]
[34]  Li JY, Chen JM, Peng ZK, 2007. Several usual gene knockout technologies. Wuyi Science Journal, 23(1): 187-190 (in Chinese) [本文引用:1]
[35]  Liang CZ, Zhang R, Guo SD, 2009. Progress of chromosome walking. Biotechnology Bulletin, 30(10): 75-83 (in Chinese) [本文引用:1]
[36]  Liu D, Gong J, Dai W, Kang X, Huang Z, Zhang HM, Liu W, Liu L, Ma J, Xia Z, Chen Y, Chen Y, Wang D, Ni P, Guo AY, Xiong X, 2012. The genome of Gand erma lucidum provide insights into triterpense biosynthesis and wood degradation. PLoS One, 7(5): e36146 [本文引用:1]
[37]  Liu F, Wang W, Xie BG, 2014. Comparison of gene expression patterns in the mycelium and primordia of Flammulina velutipes, strain 1123. Acta Edulis Fungi, 21(1): 1-7 (in Chinese) [本文引用:1]
[38]  Luo B, Chen YK, Wang Y, 2012. Methods and techniques for estimating the copy number and flanking sequences of exogenous gene in transgenic plants. Journal of Henan Normal University (Natural Science), 40(6): 111-116 (in Chinese) [本文引用:1]
[39]  Papazova N, Ghedira R, Van Glabeke S, Bartegi A, Windels P, Taverniers I, Roldan-Ruiz I, Van Bockstaele E, Milcamps A, Van Den Eede G, Depicker A, De Loose M, 2008. Stability of the T-DNA flanking regions in transgenic Arabidopsis thaliana plants under influence of abiotic stress and cultivation practice. Plant Cell Reports, 27(4): 749-757 [本文引用:2]
[40]  Park YJ, Baek JH, Lee S, Kim C, Rhee H, Kim H, Seo JS, Park HR, Yoon DE, Nam JY, Kim HI, Kim JG, Yoon H, Kang HW, Cho JY, Song ES, Sung GH, Yoo YB, Lee CS, Lee BM, Kong WS, 2014. Whole genome and global gene expression analyses of the model mushroom Flammulina velutipes reveal a high capacity for lignocellulose degradation. PLoS One, 9(4): e93560 [本文引用:1]
[41]  Pillai MM, Venkataraman GM, Kosak S, Torok-Storb B, 2008. Integration site analysis in transgenic mice by thermal asymmetric interlaced (TAIL) PCR: segregation multiple integrant fouder line and determining zygosity. Transgenic Research, 17(4): 749-754 [本文引用:2]
[42]  Qin G, Kang D, Dong Y, Shen Y, Zhang L, Deng X, Zhang Y, Li S, Chen N, Niu W, Chen C, Liu P, Chen H, Li J, Ren Y, Gu H, Deng XW, Qu LJ, Chen Z, 2003. Obtaining and analysis of flanking sequences from T-DNA transformants of Arabidopsis. Plant Science, 165(5): 941-949 [本文引用:2]
[43]  Shi LL, van Peer AF, Guo LX, Chen RL, Wang W, Yan JJ, Deng YJ, Xie BG, 2014. Agrobacterium-mediated transformation of an endogenous HMG-box transcription factor fvhom1 in Flammulina velutipes. Genomics and Applied Biology, 33(6): 1268-1274 (in Chinese) [本文引用:1]
[44]  Walden R, 2002. T-DNA tagging in a genomics era. Plant Sciences, 21(3): 143-165 [本文引用:1]
[45]  Wang K, Zhang YJ, Guan B, Guo WZ, Zhang TZ, 2007. Fluorescence in situ hybridization of bacterial artificial chromosome in cotton. Progress in Biochemistry and Biophysics, 34(11): 1216-1222 (in Chinese) [本文引用:1]

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