CN115404194A - 一株毒力致弱的牛支原体基因突变株及其应用 - Google Patents
一株毒力致弱的牛支原体基因突变株及其应用 Download PDFInfo
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Abstract
本发明公开了一株牛支原体基因突变株,命名为牛支原体(Mycoplasma bovis)T8.66,保藏在中国典型培养物保藏中心保藏,保藏编号为CCTCC NO:M 2022259,发生突变的基因是核苷酸序列如SEQ ID NO:1所示的Mbov_0725基因。T8.66突变株相比野生株和回补株对EBL和MAC‑T细胞的黏附能力下降,由于上皮细胞黏附是牛支原体毒力因素的重要表型之一,因此该毒力致弱菌株在牛支原体疫苗的制备中发挥重要作用,在牛支原体致病机制研究中也具有潜在应用价值。
Description
技术领域
本发明属于动物传染病防治技术领域,具体涉及一株牛支原体基因突变株,该菌株由于Mbov_0725基因发生突变从而对牛肺上皮细胞(EBL)和牛乳腺上皮细胞(MAC-T)的黏附能力降低,导致毒力致弱。
背景技术
牛支原体(Mycoplasma bovis)是引起牛支原体病的主要病原,也是牛呼吸道系统综合征的重要条件致病菌之一,通常牛支原体感染后主要引起的临床症状有肺炎、乳腺炎、关节炎、结膜炎、生殖系统疾病等(Perez-Casal,2020)。研究发现,牛支原体可定殖于健康牛上呼吸道而不引起任何临床症状,当发生断奶、长途运输等刺激后,导致牛只免疫力低下,与其它细菌和病毒混合感染而导致呼吸道系统综合征的发生(Calcutt et al.,2018)。目前,国内仍无有效疫苗和抗生素用于牛支原体病的防治。
研究发现,牛支原体的主要致病因素包括细胞表面可变脂蛋白,黏附和入侵宿主细胞,调控宿主免疫系统,生物膜形成,释放二级代谢产物如过氧化氢,与其它病原的协同感染等(Burki et al.,2015a)。支原体黏附于粘膜上皮细胞是其感染的第一步,不仅有利于其突破宿主细胞的屏障结构,持续增殖,向深层组织侵袭,而且可以激发宿主免疫反应引起疾病发生(Burki et al.,2015b)。目前已经报道了一些黏附相关蛋白如mbfN(Adamu etal.,2021),fructose-1.6-bisphosphate aldolase(Yu et al.,2018),NOX(Zhao et al.,2017)等,而这些蛋白的功能大多基于原核表达的重组蛋白,在牛支原体菌体水平的作用仍尚不清楚。申请人利用牛支原体转座子突变体库结合牛支原体细胞黏附模型,筛选到一株细胞黏附能力相比野生株降低的突变体T8.66,该突变株突变的基因Mbov_0725编码的蛋白是一种卤酸脱卤酶(haloacid dehalogenase,HAD)超家族成员之一,在牛支原体中具有丝氨酸/苏氨酸磷酸酶活性,参与牛支原体核苷酸代谢等过程。
发明内容
本发明的目的在于提供一株牛支原体基因突变株,该突变株呈现显著的黏附降低表型,鉴于黏附是目前牛支原体毒力反应的重要表型之一,该突变株可望在牛支原体致病、生理代谢和免疫防控领域有潜在应用前景。
为了实现本发明的目的,申请人所在的农业微生物学国家重点实验室反刍动物病原分室从牛支原体突变体库中筛选出Mbov_0725基因缺失突变株T8.66,发生突变的Mbov_0725基因其核苷酸序列如SEQ ID NO:1所示,序列长度为858bp,所编码的蛋白质序列如SEQID NO:2所示。基于两种宿主上皮细胞检测突变株/野生株/回补株与细胞的黏附能力,结果显示,相比野生株和回补株,突变株T8.66黏附宿主上皮细胞的能力显著降低。鉴于上皮细胞黏附为牛支原体毒力因素的重要表型之一,因此本发明获得的牛支原体突变株T8.66可望在牛支原体致病机制研究和免疫防制药物研制等方面具有潜在的应用价值。
具体地,本发明的技术方案如下所述:
申请人以之前从湖北省应城市某养牛场发病的黄牛的肺组织中分离得到的一株牛支原体本地分离株HB0801(Mycoplasma bovis HB0801)为亲本株,利用PEG介导的转化方法,将含有转座子的pMT85质粒转化到牛支原体中,利用庆大霉素做抗性筛选标志,对突变菌株进行筛选,成功构建了牛支原体突变体库,从该突变体库中筛选到一系列具有特定功能的突变株并已陆续申请专利。本次发明从突变体库中筛选出的一株突变株T8.66,申请人将该突变株命名为牛支原体T8.66,Mycoplasma bovis T8.66,于2022年3月15日送交中国典型培养物保藏中心保藏,保藏编号为CCTCC NO:M 2022259。利用牛肺上皮细胞(EBL)和牛乳腺上皮细胞(MAC-T)为黏附模型,检测突变株T8.66/回补株及野生株对细胞的黏附能力,证明Mbov_0725基因缺失后黏附能力显著降低。
本发明具有以下优点:
(1)本发明的突变株是发明人从牛支原体基因缺失突变体库中通过大量随机筛选而获得,不仅验证了其表型和功能,而且还对突变位点进行了鉴定,证实其发生突变的基因是Mbov_0725基因。
(2)本发明提供的T8.66突变株相比于野生株HB0801和回补株对EBL和MAC-T细胞的黏附能力下降。由于上皮细胞黏附为牛支原体毒力因素的重要表型之一,因此该毒力致弱菌株不仅在牛支原体疫苗的制备中发挥重要作用,而且还在牛支原体致病机制研究中具有潜在应用价值。
更详细的技术方案参见具体实施例。
附图说明
图1:Mbov_0725基因序列中转座子插入突变位点。图中:紫色方框所示序列为转座子与HB0801基因组连接处测序序列,所示方向为转座子相对基因组的插入方向。
图2:是缺失株T8.66和回补菌株CT8.66的Western blot检测、生长曲线和菌落形态。A.是Western blot鉴定牛支原体T8.66菌株缺失表达MbovP725的图。附图标记说明:MbovP579:表示抗牛支原体MbovP579蛋白的鼠单克隆抗体;MbovP725表示抗牛支原体MbovP725蛋白的鼠多克隆抗体。B.是牛支原体各菌株在PPLO培养基中生长曲线分析图。C:野生株及突变株/回补株中的菌落形态图。
图3:是牛支原体突变株T8.66与野生株/回补株感染牛肺上皮细胞EBL和牛乳腺上皮细胞MAC-T后黏附能力定量检测分析。附图标记说明:“*”表示P﹤0.05,“**”表示P﹤0.01,“***”表示P﹤0.001。
具体实施方式
实施例1:牛支原体Mbov_0725基因缺失/回补株构建与鉴定
1.1T8.66菌株突变基因的鉴定
利用细菌基因组提取试剂盒(购自宝生物工程大连有限公司),提取牛支原体T8.66突变体全基因组,对Tn4001转座子与牛支原体基因组连接处进行测序,将测序结果与牛支原体HB0801全基因组序列进行比对,结果显示,T8.66突变基因为Mbov_0725,转座子插入位点位于基因组857920位点后,位于Mbov_0725基因108位点后(图1)。
1.2牛支原体Mbov_0725基因缺失/回补株构建
从实验室前期构建的牛支原体突变体库中筛选出Mbov_0725基因缺失菌株T8.66,并且通过Western blot在蛋白水平上验证Mbov_0725基因缺失;回补突变体的构建如下:以牛支原体基因组为模板,扩增Mbov_0725基因,将目的片段克隆至pOH/P质粒,获得回补质粒;利用PEG8000(购自SIGMA)介导的转化方法,将构建好的重组质粒转化至Mbov_0725基因缺失菌株中,在固体培养基中培养3-7天,利用抗性挑选阳性克隆。将阳性克隆扩大培养后,提取菌体的总蛋白,利用Western blot方法验证重组菌中MbovP725蛋白的表达。在PPLO培养基中,检测突变菌株/回补菌株的生长曲线并与野生株比较,确定突变菌株/回补菌株的生长未受到影响。实验结果显示MbovP725蛋白在缺失株T8.66中不表达,在回补菌株CT8.66中表达,且缺失株T8.66和回补菌株CT8.66与野生株HB0801生长曲线和菌落形态无差异(图2)。
实施例2:牛支原体对上皮细胞的黏附能力的测定
2.1EBL细胞和MAC-T细胞的培养
EBL细胞培养于含10%热灭活胎牛血清的DMEM培养基(购自美国Hyclone公司),MAC-T细胞培养于10%热灭活胎牛血清DMEM-F12完全培养基(购自美国Hyclone公司),于37℃,5%CO2条件下培养。
2.2牛支原体各菌株的黏附检测
将生长于对数期的牛支原体各菌株进行CFU计数,取计数好的牛支原体HB0801,突变株T8.66以及回补菌株CT8.66用PBS洗涤3次,并用相应的细胞培养基进行等体积重悬,按照感染比(MOI)为1000的比例分别感染EBL和MAC-T细胞,在37℃,5%CO2条件下感染30min,60min,120min,感染后用无菌PBS洗涤三次,以洗去未黏附的牛支原体,加入1ml超纯水室温放置10min,以充分裂解细胞,然后进行CFU计数。结果显示T8.66的黏附能力较野生株和回补株均有显著降低(图3)。
综上所述,与牛支原体野生株比较,牛支原体突变株T8.66表现出MbovP725蛋白缺失,对细胞的黏附能力降低,这些特征导致该突变株可能毒力致弱,这将在牛支原体致病机理研究和免疫防制中发挥重要作用。
名词术语说明:
牛支原体Mbov_0725编码蛋白:以MbovP725表示。
牛支原体Mbov_0725基因突变株:以牛支原体T8.66表示。
牛支原体Mbov_0725基因回补株:以牛支原体CT8.66表示。
牛支原体野生株:以牛支原体HB0801表示。
牛支原体Mbov_P579编码蛋白:以MbovP579表示,该蛋白已在CN107118262A的专利文献中公开。
对序列表的说明:
SEQ ID NO:1是本发明的牛支原体蛋白基因Mbov_0725的核苷酸序列,序列长度为858bp。
SEQ ID NO:2是牛支原体蛋白基因Mbov_0725编码的蛋白质序列。
参考文献:
[1].Adamu,J.Y.,Mitiku,F.,Hartley,C.A.,Sansom,F.M.,Marenda,M.S.,Markham,P.F.,Browning,G.F.,Tivendale,K.A.,2021.Mycoplasma bovis mbfN Encodesa Novel LRR Lipoprotein That Undergoes Proteolytic Processing and Binds HostExtracellular Matrix Components.Journal of Bacteriology 203.
[2].Burki,S.,Frey,J.,Pilo,P.,2015a.Virulence,persistence anddissemination of Mycoplasma bovis.Vet Microbiol 179,15-22.
[3].Burki,S.,Gaschen,V.,Stoffel,M.H.,Stojiljkovic,A.,Frey,J.,Kuehni-Boghenbor,K.,Pilo,P.,2015b.Invasion and persistence of Mycoplasma bovis inembryonic calf turbinate cells.Vet Res 46,53.
[4].Calcutt,M.J.,Lysnyansky,I.,Sachse,K.,Fox,L.K.,Nicholas,R.A.J.,Ayling,R.D.,2018.Gap analysis of Mycoplasma bovis disease,diagnosis andcontrol:An aid to identify future development requirements.Transbound EmergDis 65 Suppl 1,91-109.
[5].Perez-Casal,J.,2020.Pathogenesis and Virulence of Mycoplasmabovis.Vet Clin North Am Food Anim Pract 36,269-278.
[6].Yu,Y.,Liu,M.,Hua,L.,Qiu,M.,Zhang,W.,Wei,Y.,Gan,Y.,Feng,Z.,Shao,G.,Xiong,Q.,2018.Fructose-1,6-bisphosphate aldolase encoded by a core gene ofMycoplasma hyopneumoniae contributes to host cell adhesion.Vet Res 49,114.
[7].Zhao,G.,Zhang,H.,Chen,X.,Zhu,X.,Guo,Y.,He,C.,Anwar Khan,F.,Chen,Y.,Hu,C.,Chen,H.,Guo,A.,2017.Mycoplasma bovis NADH oxidase functions as botha NADH oxidizing and O2 reducing enzyme and an adhesin.Sci Rep 7,44.
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<120> 一株毒力致弱的牛支原体基因突变株及其应用
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Claims (3)
1.一株牛支原体基因突变株,命名为牛支原体(Mycoplasma bovis)T8.66,保藏在中国典型培养物保藏中心保藏,保藏编号为CCTCC NO:M 2022259。
2.如权利要求1所述的牛支原体基因突变株,该菌株发生突变的基因是核苷酸序列如SEQ ID NO:1所示的Mbov_0725基因。
3.权利要求1所述的牛支原体基因突变株在制备牛支原体疫苗中的应用,该菌株对牛肺上皮细胞(EBL)和牛乳腺上皮细胞(MAC-T)的黏附能力降低,导致毒力致弱。
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---|---|---|---|---|
CN109837226A (zh) * | 2019-02-21 | 2019-06-04 | 华中农业大学 | 黏附能力降低的牛支原体基因突变株及黏附蛋白 |
CN111635880A (zh) * | 2020-05-06 | 2020-09-08 | 华中农业大学 | 一种牛支原体VspX基因突变株及其构建方法和应用 |
CN111676181A (zh) * | 2020-05-09 | 2020-09-18 | 华中农业大学 | 牛支原体Mbov_0145基因突变株及其应用 |
CN111705013A (zh) * | 2020-06-02 | 2020-09-25 | 华中农业大学 | 一种牛支原体Mbov_0570基因突变株及其应用 |
CN111748507A (zh) * | 2020-06-02 | 2020-10-09 | 华中农业大学 | 一种牛支原体Mbov_0475基因突变株及其应用 |
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Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109837226A (zh) * | 2019-02-21 | 2019-06-04 | 华中农业大学 | 黏附能力降低的牛支原体基因突变株及黏附蛋白 |
CN111235082A (zh) * | 2019-02-21 | 2020-06-05 | 华中农业大学 | 黏附能力降低的牛支原体基因缺失的突变株 |
CN111635880A (zh) * | 2020-05-06 | 2020-09-08 | 华中农业大学 | 一种牛支原体VspX基因突变株及其构建方法和应用 |
CN111676181A (zh) * | 2020-05-09 | 2020-09-18 | 华中农业大学 | 牛支原体Mbov_0145基因突变株及其应用 |
CN111705013A (zh) * | 2020-06-02 | 2020-09-25 | 华中农业大学 | 一种牛支原体Mbov_0570基因突变株及其应用 |
CN111748507A (zh) * | 2020-06-02 | 2020-10-09 | 华中农业大学 | 一种牛支原体Mbov_0475基因突变株及其应用 |
Non-Patent Citations (1)
Title |
---|
李明霞的: "牛支原体黏附和侵入宿主细胞的研究进展", 中国兽医学报, vol. 38, no. 7 * |
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