茅光耀:小管福寿螺Cu2+抗性相关基因筛选及其抗性机制初探论文

茅光耀:小管福寿螺Cu2+抗性相关基因筛选及其抗性机制初探论文

本文主要研究内容

作者茅光耀(2019)在《小管福寿螺Cu2+抗性相关基因筛选及其抗性机制初探》一文中研究指出:目的:小管福寿螺(Pomaceacanaliculata是一种典型的入侵性食草生物,更是一种重要热带病传播相关媒介生物。其在广州管圆线虫病的发生和传播过程中起到了非常重要的作用,对我国居民健康构成了巨大的威胁。目前,我国小管福寿螺的防制效果不太理想。该物种易于入侵、难于防制的重要原因之一是其极强的环境适应能力。开展对小管福寿螺的环境适应性机制的研究,有助于深入了解该重要热带病传播相关入侵媒介生物的生物学特性,对于制定和完善我国今后的小管福寿螺防制策略具有重要意义。野外调查显示,小管福寿螺对重金属污染严重水体适应性较强。相比于Fe,Cd等重金属,重金属Cu在电镀冶金、机械制造、消毒剂及农药生产等诸多方面有着更为广泛的应用,更易进入水体造成污染,水体中的Cu主要以CLu2+形式存在,其生物毒性显著,污染情况日益严峻。研究表明,小管福寿螺的Cu2+耐受能力显著高于其他淡水螺群。但是对小管福寿螺Ca2+适应性较强这一现象的研究,目前仅限于小管福寿螺自身趋避行为和病理生理学变化,相关功能基因的研究尚未见有涉及。因此,本研究着眼于筛选出一批小管福寿螺Cu2+抗性相关功能基因,为深入研究小管福寿螺Cu2+胁迫适应的分子机制提供有价值的候选基因。此外,本研究对候选基因中的金属硫蛋白基因(PcMT)和谷胱甘肽硫转移酶基因(PcGST1和PcGST2)的基本特征、与其他物种同源基因的遗传进化关系、组织表达本底水平、Cu2+胁迫前后表达量变化情况和Cu2+抗性功能进行分析,并构建原核表达载体,获得纯化的PcMT和PcGST2蛋白,初步探索小管福寿螺可能的Cu2+耐受机制。方法:①在Cu2+胁迫下小管福寿螺血液转录组测序获得的结果基础上,筛选出一批Cu2+抗性相关功能基因,对这些基因的可靠性通过实时荧光定量PCR方法验证。②选择其中的PcMT、PcGST1和PcGST2基因开展进一步研究,通过PCR方法扩增基因编码区,利用生物学信息学方法分析基因基本特征,利用MEGA 7.0软件构建进化树分析与其他物种同源基因的遗传进化关系,利用实时荧光定量PCR方法分析组织(肝脏、肾脏、鳃、血液和肠)基因的本底表达水平和Cu2+(50、100、150 μg/L)胁迫下(0、1、7、14d)组织中(肝脏、肾脏、鳃)基因的表达量变化情况。③选择pET28a为表达载体,构建pET28a-PcMT和pET28a-PcGST2质粒,利用IPTG诱导PcMT和PcGST2蛋白在大肠杆菌BL21(DE3)中表达,利用Ni-NDA亲和层析法纯化PcMT和PcGST2蛋白。分析OD600值变化,比较含pET28a-PcMT和pET28a-PcGST2质粒的转化菌株与含pET28a空载体的非转化菌株在Cu2+胁迫下生长情况,初步验证PcMT和PcGST2基因Cu2+抗性功能。结果:小管福寿螺体内金属硫蛋白、热休克蛋白、谷胱甘肽硫转移酶、复肌球蛋白和天冬氨酸氧化酶等17个基因的差异表达与Cu2+胁迫存在一定的相关性。②研究的PcMT基因编码区全长272bp,共编码87个氨基酸,不含芳香族氨基酸和组氨酸,其末端序列CQCGKGCTGADSCKCDRKCSCK与软体动物MT特征保守序列CXCXXXCTGXXXCXCXXXCXCK完全一致,属于软体动物金属硫蛋白家族2:亚家族mog:腹足纲金属硫蛋白。PcMT蛋白含有较高的Cys残基(22个,25.3%)和较多的巯基金属簇的特征保守结构Cys-X(1-3)-Cys(17个),可能具有很强的金属结合能力,经氨基酸序列对比,PcMT与同属另一种福寿螺Pomacea bridgesi的金属硫蛋白同源性最高,为93%。③以未经Cu2+胁迫的小管福寿螺肝脏组织样本第一次实验获得的结果为对照1.0±0.0,发现PcMT基因的表达具有组织差异性,在小管福寿螺各组织中的表达量分别为:鳃2.4±0.4>肝1.0±0.1>肠0.9±0.1>肾脏0.7±0.1>血液0.5±0.09。④小管福寿螺PcT基因对环境中Cu2+敏感,以未经Cu2+胁迫的小管福寿螺对应组织样本第一次实验结果作为对照1.0±0.0,50μg/L即可诱导该基因在肝脏、鳃和肾组织中表达,且诱导表达的效果与胁迫时间呈较好的正相关,14d时基因表达量分别为2.03±0.3,3.4±0.4,2.8±0.6。100μg/LCu2+胁迫下,小管福寿螺肝脏组织中PcMT基因表达量与胁迫时长呈正相关,14d时基因表达量为2.3±0.3。而在肾组织和鳃组织中,表达量与胁迫时间呈现倒U型的特征,峰值均出现在第7d,分别为5.2±0.6和8.5±1.1。150 μ g/L Cu2t胁迫下,小管福寿螺肝脏、鳃和肾组织中PcMT基因的表达量与胁迫时间均呈现倒U型的特征,肝脏组织峰值出现在第7d,为3.3±0.8,鳃和肾组织峰值出现在第1d,分别为7.001±0.4、9.1±0.8。⑤研究的PcGST1和PcGST2基因编码区全长分别为591bp和603bp,分别编码196和200个氨基酸,末端分别含有GST-C-3和GST-C-σ超家族结构域,可与重金属胁迫产生的活性氧自由基等有害亲电子底物结合,两者N末端均为GST-N-σ结构域超家族,可与还原型谷胱甘肽结合。PcGST1和PcGST2与同腹足纲的加州海兔(Aplysia californic)和光滑双脐螺(Biomphalariaglabrata)亲缘关系较近,聚为一支,而与双壳纲软体动物夷虾扇贝(Mizuhopecten yessoensis)、栉孔扇贝(Azumapectenfarreri)和海湾扇(Argopecten irradianm)等的 GSTs蛋白亲缘关系较远,分支较长,σ型GST蛋白可能在软体动物腹足纲和双壳纲形成过程中出现了一定程度的分化。⑥同亚型不同谷胱甘肽硫转移酶基因之间组织本底表达水平不完全相同,以未经Cu2+胁迫的小管福寿螺肝脏组织样本第一次实验结果作为对照1.0±0.0,PcGST1基因本底表达水平为:血液8.5±0.3>鳃1.4±0.2>肝脏1.1±0.2>肠1.03±0.1>肾脏0.4±0.08,PcGST2基因本底表达水平为:血液5.8±0.9>肠 1.6±0.4>肝脏0.8±0.1>鳃0.2±0.02>肾脏0.2±0.01。⑦同亚型不同的谷胱甘肽硫转移酶基因之间Cu2+胁迫下表达模式不完全相同,以未经Cu2+胁迫的小管福寿螺对应组织样本第一次实验结果作为对照1.0±0.0,Cu2+胁迫对PcGST1基因表达的影响为:肝组织中,50μg/L和100μg/LCu2+胁迫下PcGST1基因表达量0-14d持续上升,14d时表达量分别为2.1±0.1和5.2±0.7,而150μg/Lu2+胁迫下,1d达到峰值,为11.3±1.8。鳃组织中,50μg/L浓度Cu2+胁迫下基因表达量变化不明显,而150 u g/L浓度Cu2+胁迫时1d即可达到表达量峰值,为15.8±8.3。肾组织中50μg/L、100μg/L和1500μg/L浓度均可诱导表达,但50 μμg/L诱导期较短为7d,表达峰值较低,为3.1±0.3,150 μg/L诱导期较长为14d,但表达峰值较高,为5.6±2.6。⑧以未经Cu2+胁迫的小管福寿螺对应组织样本第一次实验结果作为对照1.0±0.0,Cu2胁迫对PcGST2基因表达的影响为:肝组织中,50μg/LCu2+胁迫下0-14d基因表达量持续上升,14d时表达量为4.9±0.5,100、150 μg/LCu2+胁迫下,PcGST2基因的相对表达量在0-7d呈上升趋势,7d时表达量分别为13.01士3.03和12.2±2.2,随后呈下降趋势。鳃组织中,50、100 ug/LCu2+胁迫下,PcGST2基因的相对表达量的峰值均于7d达到峰值,分别为5.3±0.7、23.3±16.5,150 μg/LCu2+胁迫下,PcGST2基因的相对表达量于1 d达到峰值,为9.8±3.3。肾组织中,50 μg/LCu2+胁迫下,PcGST2基因的相对表达量随时间变化不显著,100 μg/LCu2+胁迫下,PcGST2基因的相对表达量前期随时间变化不显著,约于14 d时出现显著增加,表达量为3.9士1.04;150 μg/LCu2+胁迫下,PcGST2基因的相对表达量呈持续上升趋势,14d表达量为18.02±0.8。⑨成功构建了含pET28a-PcMT和pET28a-PcGST2质粒的BL21(DE3)大肠杆菌,在1mmol/LIPTG、20℃诱导下,分别可表达出17 000的PcMT蛋白和27 000的PcGST2蛋白,经Ni-NDA亲和层析纯化,最终获得了PcMT和PcGST2蛋白。⑩含pET28a-PcMT、pET28a-PcGST2质粒的转化菌株与含pET28a空载体的非转化菌株在正常的ILB液体培养基中,生长曲线接近,差异无统计学意义。而在含0.2mmol/LCuSO4的LB液体培养基中,转化菌株Cu2+的耐性较强,生长曲线优于非转化菌株,差异有统计学意义。结论:①小管福寿螺Cu2+适应的分子机制非常复杂,涉及到众多基因的协调参与,经实时荧光定量PCR验证的17个差异表达基因可靠性较高,可作为深入研究小管福寿螺Cu2+胁迫适应分子机理的候选基因。②Cu2+胁迫可以显著诱导小管福寿螺体内金属硫蛋白基因和谷胱甘肽硫转移酶基因的表达,金属硫蛋白基因和谷胱甘肽硫转移酶基因可能在小管福寿螺Cu2+胁迫适应过程中发挥了一定的作用。③小管福寿螺金属硫蛋白基因和谷胱甘肽硫转移酶基因可赋予转化菌株一定的Cu2+抗性能力,具有Cu2+抗性功能,这为下一步获得“金属硫蛋白基因和谷胱甘肽硫转移酶基因可提高小管福寿螺Cu2+耐受能力”直接证据奠定了基础。

Abstract

mu de :xiao guan fu shou luo (Pomaceacanaliculatashi yi chong dian xing de ru qin xing shi cao sheng wu ,geng shi yi chong chong yao re dai bing chuan bo xiang guan mei jie sheng wu 。ji zai an zhou guan yuan xian chong bing de fa sheng he chuan bo guo cheng zhong qi dao le fei chang chong yao de zuo yong ,dui wo guo ju min jian kang gou cheng le ju da de wei xie 。mu qian ,wo guo xiao guan fu shou luo de fang zhi xiao guo bu tai li xiang 。gai wu chong yi yu ru qin 、nan yu fang zhi de chong yao yuan yin zhi yi shi ji ji jiang de huan jing kuo ying neng li 。kai zhan dui xiao guan fu shou luo de huan jing kuo ying xing ji zhi de yan jiu ,you zhu yu shen ru le jie gai chong yao re dai bing chuan bo xiang guan ru qin mei jie sheng wu de sheng wu xue te xing ,dui yu zhi ding he wan shan wo guo jin hou de xiao guan fu shou luo fang zhi ce lve ju you chong yao yi yi 。ye wai diao cha xian shi ,xiao guan fu shou luo dui chong jin shu wu ran yan chong shui ti kuo ying xing jiao jiang 。xiang bi yu Fe,Cddeng chong jin shu ,chong jin shu Cuzai dian du ye jin 、ji xie zhi zao 、xiao du ji ji nong yao sheng chan deng zhu duo fang mian you zhao geng wei an fan de ying yong ,geng yi jin ru shui ti zao cheng wu ran ,shui ti zhong de Cuzhu yao yi CLu2+xing shi cun zai ,ji sheng wu du xing xian zhe ,wu ran qing kuang ri yi yan jun 。yan jiu biao ming ,xiao guan fu shou luo de Cu2+nai shou neng li xian zhe gao yu ji ta dan shui luo qun 。dan shi dui xiao guan fu shou luo Ca2+kuo ying xing jiao jiang zhe yi xian xiang de yan jiu ,mu qian jin xian yu xiao guan fu shou luo zi shen qu bi hang wei he bing li sheng li xue bian hua ,xiang guan gong neng ji yin de yan jiu shang wei jian you she ji 。yin ci ,ben yan jiu zhao yan yu shai shua chu yi pi xiao guan fu shou luo Cu2+kang xing xiang guan gong neng ji yin ,wei shen ru yan jiu xiao guan fu shou luo Cu2+xie pai kuo ying de fen zi ji zhi di gong you jia zhi de hou shua ji yin 。ci wai ,ben yan jiu dui hou shua ji yin zhong de jin shu liu dan bai ji yin (PcMT)he gu guang gan tai liu zhuai yi mei ji yin (PcGST1he PcGST2)de ji ben te zheng 、yu ji ta wu chong tong yuan ji yin de wei chuan jin hua guan ji 、zu zhi biao da ben de shui ping 、Cu2+xie pai qian hou biao da liang bian hua qing kuang he Cu2+kang xing gong neng jin hang fen xi ,bing gou jian yuan he biao da zai ti ,huo de chun hua de PcMThe PcGST2dan bai ,chu bu tan suo xiao guan fu shou luo ke neng de Cu2+nai shou ji zhi 。fang fa :①zai Cu2+xie pai xia xiao guan fu shou luo xie ye zhuai lu zu ce xu huo de de jie guo ji chu shang ,shai shua chu yi pi Cu2+kang xing xiang guan gong neng ji yin ,dui zhe xie ji yin de ke kao xing tong guo shi shi ying guang ding liang PCRfang fa yan zheng 。②shua ze ji zhong de PcMT、PcGST1he PcGST2ji yin kai zhan jin yi bu yan jiu ,tong guo PCRfang fa kuo zeng ji yin bian ma ou ,li yong sheng wu xue xin xi xue fang fa fen xi ji yin ji ben te zheng ,li yong MEGA 7.0ruan jian gou jian jin hua shu fen xi yu ji ta wu chong tong yuan ji yin de wei chuan jin hua guan ji ,li yong shi shi ying guang ding liang PCRfang fa fen xi zu zhi (gan zang 、shen zang 、sai 、xie ye he chang )ji yin de ben de biao da shui ping he Cu2+(50、100、150 μg/L)xie pai xia (0、1、7、14d)zu zhi zhong (gan zang 、shen zang 、sai )ji yin de biao da liang bian hua qing kuang 。③shua ze pET28awei biao da zai ti ,gou jian pET28a-PcMThe pET28a-PcGST2zhi li ,li yong IPTGyou dao PcMThe PcGST2dan bai zai da chang gan jun BL21(DE3)zhong biao da ,li yong Ni-NDAqin he ceng xi fa chun hua PcMThe PcGST2dan bai 。fen xi OD600zhi bian hua ,bi jiao han pET28a-PcMThe pET28a-PcGST2zhi li de zhuai hua jun zhu yu han pET28akong zai ti de fei zhuai hua jun zhu zai Cu2+xie pai xia sheng chang qing kuang ,chu bu yan zheng PcMThe PcGST2ji yin Cu2+kang xing gong neng 。jie guo :xiao guan fu shou luo ti nei jin shu liu dan bai 、re xiu ke dan bai 、gu guang gan tai liu zhuai yi mei 、fu ji qiu dan bai he tian dong an suan yang hua mei deng 17ge ji yin de cha yi biao da yu Cu2+xie pai cun zai yi ding de xiang guan xing 。②yan jiu de PcMTji yin bian ma ou quan chang 272bp,gong bian ma 87ge an ji suan ,bu han fang xiang zu an ji suan he zu an suan ,ji mo duan xu lie CQCGKGCTGADSCKCDRKCSCKyu ruan ti dong wu MTte zheng bao shou xu lie CXCXXXCTGXXXCXCXXXCXCKwan quan yi zhi ,shu yu ruan ti dong wu jin shu liu dan bai jia zu 2:ya jia zu mog:fu zu gang jin shu liu dan bai 。PcMTdan bai han you jiao gao de Cyscan ji (22ge ,25.3%)he jiao duo de qiu ji jin shu cu de te zheng bao shou jie gou Cys-X(1-3)-Cys(17ge ),ke neng ju you hen jiang de jin shu jie ge neng li ,jing an ji suan xu lie dui bi ,PcMTyu tong shu ling yi chong fu shou luo Pomacea bridgeside jin shu liu dan bai tong yuan xing zui gao ,wei 93%。③yi wei jing Cu2+xie pai de xiao guan fu shou luo gan zang zu zhi yang ben di yi ci shi yan huo de de jie guo wei dui zhao 1.0±0.0,fa xian PcMTji yin de biao da ju you zu zhi cha yi xing ,zai xiao guan fu shou luo ge zu zhi zhong de biao da liang fen bie wei :sai 2.4±0.4>gan 1.0±0.1>chang 0.9±0.1>shen zang 0.7±0.1>xie ye 0.5±0.09。④xiao guan fu shou luo PcTji yin dui huan jing zhong Cu2+min gan ,yi wei jing Cu2+xie pai de xiao guan fu shou luo dui ying zu zhi yang ben di yi ci shi yan jie guo zuo wei dui zhao 1.0±0.0,50μg/Lji ke you dao gai ji yin zai gan zang 、sai he shen zu zhi zhong biao da ,ju you dao biao da de xiao guo yu xie pai shi jian cheng jiao hao de zheng xiang guan ,14dshi ji yin biao da liang fen bie wei 2.03±0.3,3.4±0.4,2.8±0.6。100μg/LCu2+xie pai xia ,xiao guan fu shou luo gan zang zu zhi zhong PcMTji yin biao da liang yu xie pai shi chang cheng zheng xiang guan ,14dshi ji yin biao da liang wei 2.3±0.3。er zai shen zu zhi he sai zu zhi zhong ,biao da liang yu xie pai shi jian cheng xian dao Uxing de te zheng ,feng zhi jun chu xian zai di 7d,fen bie wei 5.2±0.6he 8.5±1.1。150 μ g/L Cu2txie pai xia ,xiao guan fu shou luo gan zang 、sai he shen zu zhi zhong PcMTji yin de biao da liang yu xie pai shi jian jun cheng xian dao Uxing de te zheng ,gan zang zu zhi feng zhi chu xian zai di 7d,wei 3.3±0.8,sai he shen zu zhi feng zhi chu xian zai di 1d,fen bie wei 7.001±0.4、9.1±0.8。⑤yan jiu de PcGST1he PcGST2ji yin bian ma ou quan chang fen bie wei 591bphe 603bp,fen bie bian ma 196he 200ge an ji suan ,mo duan fen bie han you GST-C-3he GST-C-σchao jia zu jie gou yu ,ke yu chong jin shu xie pai chan sheng de huo xing yang zi you ji deng you hai qin dian zi de wu jie ge ,liang zhe Nmo duan jun wei GST-N-σjie gou yu chao jia zu ,ke yu hai yuan xing gu guang gan tai jie ge 。PcGST1he PcGST2yu tong fu zu gang de jia zhou hai tu (Aplysia californic)he guang hua shuang qi luo (Biomphalariaglabrata)qin yuan guan ji jiao jin ,ju wei yi zhi ,er yu shuang ke gang ruan ti dong wu yi ha shan bei (Mizuhopecten yessoensis)、zhi kong shan bei (Azumapectenfarreri)he hai wan shan (Argopecten irradianm)deng de GSTsdan bai qin yuan guan ji jiao yuan ,fen zhi jiao chang ,σxing GSTdan bai ke neng zai ruan ti dong wu fu zu gang he shuang ke gang xing cheng guo cheng zhong chu xian le yi ding cheng du de fen hua 。⑥tong ya xing bu tong gu guang gan tai liu zhuai yi mei ji yin zhi jian zu zhi ben de biao da shui ping bu wan quan xiang tong ,yi wei jing Cu2+xie pai de xiao guan fu shou luo gan zang zu zhi yang ben di yi ci shi yan jie guo zuo wei dui zhao 1.0±0.0,PcGST1ji yin ben de biao da shui ping wei :xie ye 8.5±0.3>sai 1.4±0.2>gan zang 1.1±0.2>chang 1.03±0.1>shen zang 0.4±0.08,PcGST2ji yin ben de biao da shui ping wei :xie ye 5.8±0.9>chang 1.6±0.4>gan zang 0.8±0.1>sai 0.2±0.02>shen zang 0.2±0.01。⑦tong ya xing bu tong de gu guang gan tai liu zhuai yi mei ji yin zhi jian Cu2+xie pai xia biao da mo shi bu wan quan xiang tong ,yi wei jing Cu2+xie pai de xiao guan fu shou luo dui ying zu zhi yang ben di yi ci shi yan jie guo zuo wei dui zhao 1.0±0.0,Cu2+xie pai dui PcGST1ji yin biao da de ying xiang wei :gan zu zhi zhong ,50μg/Lhe 100μg/LCu2+xie pai xia PcGST1ji yin biao da liang 0-14dchi xu shang sheng ,14dshi biao da liang fen bie wei 2.1±0.1he 5.2±0.7,er 150μg/Lu2+xie pai xia ,1dda dao feng zhi ,wei 11.3±1.8。sai zu zhi zhong ,50μg/Lnong du Cu2+xie pai xia ji yin biao da liang bian hua bu ming xian ,er 150 u g/Lnong du Cu2+xie pai shi 1dji ke da dao biao da liang feng zhi ,wei 15.8±8.3。shen zu zhi zhong 50μg/L、100μg/Lhe 1500μg/Lnong du jun ke you dao biao da ,dan 50 μμg/Lyou dao ji jiao duan wei 7d,biao da feng zhi jiao di ,wei 3.1±0.3,150 μg/Lyou dao ji jiao chang wei 14d,dan biao da feng zhi jiao gao ,wei 5.6±2.6。⑧yi wei jing Cu2+xie pai de xiao guan fu shou luo dui ying zu zhi yang ben di yi ci shi yan jie guo zuo wei dui zhao 1.0±0.0,Cu2xie pai dui PcGST2ji yin biao da de ying xiang wei :gan zu zhi zhong ,50μg/LCu2+xie pai xia 0-14dji yin biao da liang chi xu shang sheng ,14dshi biao da liang wei 4.9±0.5,100、150 μg/LCu2+xie pai xia ,PcGST2ji yin de xiang dui biao da liang zai 0-7dcheng shang sheng qu shi ,7dshi biao da liang fen bie wei 13.01shi 3.03he 12.2±2.2,sui hou cheng xia jiang qu shi 。sai zu zhi zhong ,50、100 ug/LCu2+xie pai xia ,PcGST2ji yin de xiang dui biao da liang de feng zhi jun yu 7dda dao feng zhi ,fen bie wei 5.3±0.7、23.3±16.5,150 μg/LCu2+xie pai xia ,PcGST2ji yin de xiang dui biao da liang yu 1 dda dao feng zhi ,wei 9.8±3.3。shen zu zhi zhong ,50 μg/LCu2+xie pai xia ,PcGST2ji yin de xiang dui biao da liang sui shi jian bian hua bu xian zhe ,100 μg/LCu2+xie pai xia ,PcGST2ji yin de xiang dui biao da liang qian ji sui shi jian bian hua bu xian zhe ,yao yu 14 dshi chu xian xian zhe zeng jia ,biao da liang wei 3.9shi 1.04;150 μg/LCu2+xie pai xia ,PcGST2ji yin de xiang dui biao da liang cheng chi xu shang sheng qu shi ,14dbiao da liang wei 18.02±0.8。⑨cheng gong gou jian le han pET28a-PcMThe pET28a-PcGST2zhi li de BL21(DE3)da chang gan jun ,zai 1mmol/LIPTG、20℃you dao xia ,fen bie ke biao da chu 17 000de PcMTdan bai he 27 000de PcGST2dan bai ,jing Ni-NDAqin he ceng xi chun hua ,zui zhong huo de le PcMThe PcGST2dan bai 。⑩han pET28a-PcMT、pET28a-PcGST2zhi li de zhuai hua jun zhu yu han pET28akong zai ti de fei zhuai hua jun zhu zai zheng chang de ILBye ti pei yang ji zhong ,sheng chang qu xian jie jin ,cha yi mo tong ji xue yi yi 。er zai han 0.2mmol/LCuSO4de LBye ti pei yang ji zhong ,zhuai hua jun zhu Cu2+de nai xing jiao jiang ,sheng chang qu xian you yu fei zhuai hua jun zhu ,cha yi you tong ji xue yi yi 。jie lun :①xiao guan fu shou luo Cu2+kuo ying de fen zi ji zhi fei chang fu za ,she ji dao zhong duo ji yin de xie diao can yu ,jing shi shi ying guang ding liang PCRyan zheng de 17ge cha yi biao da ji yin ke kao xing jiao gao ,ke zuo wei shen ru yan jiu xiao guan fu shou luo Cu2+xie pai kuo ying fen zi ji li de hou shua ji yin 。②Cu2+xie pai ke yi xian zhe you dao xiao guan fu shou luo ti nei jin shu liu dan bai ji yin he gu guang gan tai liu zhuai yi mei ji yin de biao da ,jin shu liu dan bai ji yin he gu guang gan tai liu zhuai yi mei ji yin ke neng zai xiao guan fu shou luo Cu2+xie pai kuo ying guo cheng zhong fa hui le yi ding de zuo yong 。③xiao guan fu shou luo jin shu liu dan bai ji yin he gu guang gan tai liu zhuai yi mei ji yin ke fu yu zhuai hua jun zhu yi ding de Cu2+kang xing neng li ,ju you Cu2+kang xing gong neng ,zhe wei xia yi bu huo de “jin shu liu dan bai ji yin he gu guang gan tai liu zhuai yi mei ji yin ke di gao xiao guan fu shou luo Cu2+nai shou neng li ”zhi jie zheng ju dian ding le ji chu 。

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论文作者分别是来自中国疾病预防控制中心的茅光耀,发表于刊物中国疾病预防控制中心2019-09-19论文,是一篇关于小管福寿螺论文,抗性相关基因论文,金属硫蛋白论文,谷胱甘肽硫转移酶论文,表达特征分析论文,中国疾病预防控制中心2019-09-19论文的文章。本文可供学术参考使用,各位学者可以免费参考阅读下载,文章观点不代表本站观点,资料来自中国疾病预防控制中心2019-09-19论文网站,若本站收录的文献无意侵犯了您的著作版权,请联系我们删除。

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茅光耀:小管福寿螺Cu2+抗性相关基因筛选及其抗性机制初探论文
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