CN114410812B - 一种鉴别鸡群对沙门氏菌易感性的snp位点、筛选及应用 - Google Patents
一种鉴别鸡群对沙门氏菌易感性的snp位点、筛选及应用 Download PDFInfo
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Abstract
本发明公开了一种鉴别鸡群对沙门氏菌易感性的SNP位点，所述SNP位点为TLR1A基因第2443位T/C变异和TLR4基因第1147位T/C变异；其中，第2443位T/C变异所在的特征序列为：TATAACCCAA[T/C]GTCTGATTAA，如SEQ ID NO.1所示；第1147位T/C变异所在的特征序列为：GAGTCTAACT[T/C]ACCTGGAGGT，如SEQ ID NO.2所示。
Description
技术领域
本发明涉及CAPs遗传标记技术领域，更具体地说是涉及一种鉴别鸡群对沙门氏菌易感性的SNP位点、筛选及应用。
背景技术
沙门氏菌是一种无芽孢、无荚膜的革兰氏阴性肠道杆菌，在自然界中广泛分布，是人畜获得性食源性细菌性肠胃炎的首要病原菌。禽沙门氏菌病根据感染病原不同可分为三类：鸡白痢、禽伤寒和禽副伤寒。鸡白痢常见于雏鸡，病雏表现为精神萎顿、缩头、翅下垂，拉白色浆糊状稀粪；禽伤寒主要危害3月龄以上的成年鸡，以肝、脾肿大，肝呈黄绿色或古铜色为特征；禽副伤寒可造成血分病，致使五脏及肠胃的生理功能异常，引起气虚、机体衰弱，死亡率高达50％。禽沙门氏菌病传播途径多为蛋垂直传播，也可通过接触病鸡或污染的饲料、饮水等经消化道水平传播。虽成年鸡感染沙门氏菌一般不表现明显症状，呈隐形带菌传播，但却是鸡沙门氏菌病流行和人类食物的主要污染源。
基于鸡TLR4、TLR1A在相应组织中的表达，在鸡基因组的位置和结构以及在先天免疫反应中的作用，使得它们成为研究沙门氏菌抵抗力理想的候选基因。具有新功能的新基因的出现是生物进化过程中的重要事件。复制被认为是产生新基因的最重要途径，而正选择促进了新基因的出现。基因的功能要通过蛋白来行使，基因的变异也是通过改变蛋白结构而发挥作用。研究TLR4和TLR1A在阳性和阴性群体间的差异能帮助科研人员了解沙门氏菌抗性的遗传基础，进而筛选能够影响抗性的位点和基因型。鸡对肠炎沙门氏菌感染的表达调控研究进展，将肠炎沙门氏菌感染小鸡，在感染不同天数后取盲肠样品分析，发现盲肠组织TLR1A的表达量显著上调。
目前对于禽沙门氏菌感染的治疗措施还是以抗生素和疫苗为主。抗生素只能抑制和杀灭机体内的沙门氏菌，不能防止病菌在鸡群中的传播。注射疫苗存在一定的安全隐患，可能毒力返强造成新疫情的发生。抗生素和疫苗等方式无法对禽沙门氏菌病进行根治。相较而言，抗病育种可能对提高畜禽抗病能力更为有效。
因此，如何筛选鸡群对沙门氏菌易感性的SNP位点，并将其应用于筛选抗沙门氏菌的畜禽中是本领域技术人员亟需解决的问题。
发明内容
有鉴于此，本发明利用酶切位点PCR-RFLP检测技术与抗病力显著关联的SNP，对SNP与坝上长尾鸡自然感染率的关联关系，筛选沙门氏菌抵抗力和易感性特定基因型，从而改变育种过程中难以对抗病力进行选择提高的情况。
为了实现上述目的，本发明采用如下技术方案：
一种鉴别鸡群对沙门氏菌易感性的SNP位点，所述SNP位点为TLR1A基因第2443位T/C变异和TLR4基因第1147位T/C变异；其中，第2443位T/C变异所在的特征序列为：TATAACCCAA[T/C]GTCTGATTAA，如SEQ ID NO.1所示；第1147位T/C变异所在的特征序列为：GAGTCTAACT[T/C]ACCTGGAGGT，如SEQ ID NO.2所示。
一种鉴别鸡群对沙门氏菌易感性的SNP位点的筛选方法，包括下述过程：
1)变异位点筛选和确定：用PAML程序和Datamonkey在线网站筛选正选择位点，找出与Ensembl上该基因碱基突变重合的位点。
2)样品采集：用2.5ml注射器采静脉血2ml，制作诊断鸡白痢鸡伤寒的抗原检测采集样品；
3)设计引物及酶切分型：利用酚-氯仿抽提法提DNA，选用合适的酶，设计引物，并进行酶切分型；
4)关联分析：运用逻辑回归方法，分析单个SNP和沙门氏菌易感性的相关性。
一种扩增所述SNP位点的引物，包括下述序列：
TLR1A-815F：5'-GTGGTGTCACTACGAGCTGTACTTT-3'，如SEQ ID NO.3所示；
TLR1A-815R：5'-AGATAGCCAAGTCACTTAATCAGCT-3'，如SEQ ID NO.4所示；
TLR4-383F：5'-TTCGGTTGGTGGACCTGAATCT-3'，如SEQ ID NO.5所示；
TLR4-383R：5'-CTCAAATCTACAACCTCCAGGT-3'，如SEQ ID NO.6所示。
所述的SNP位点在鉴别对沙门氏菌易感性鸡群中的应用。
一种鉴定对沙门氏菌易感性鸡群的方法，包括下述步骤：
1)提取待测鸡基因组DNA，以其为模板，利用本发明的上述特异性引物对进行PCR扩增反应，获得扩增产物片段。
2)对扩增产物酶切，检测本发明所述分子标记的基因型，酶切产物中对应基因组版本Galgal4:chr4:6933479位置，T/C型和T/T型对C/C型为沙门氏菌易感型。Galgal4:chr17:3569520位置，C/C型对T/T型为沙门氏菌易感型。
经由上述的技术方案可知，与现有技术相比，本发明公开了利用TLR1A、TLR4基因CAPs遗传标记提高鸡群沙门氏菌抵抗力的方法、引物，提供了TLR1A和TLR4基因的酶切位点PCR-RFLP标记。本发明首次采用CAPs技术，在TLR1A、TLR4基因变异与鸡沙门氏菌病易感性关联分析的基础上，通过SNP位点与基因型遗传变异分析，揭示了TLR1A、TLR4基因为鸡抵抗沙门氏菌病的功能基因，建立了高效率的2个基因的CAPs检测标记，对基因纯化和抗病育种具有积极的指导意义。该技术操作简单，准确性高，将本遗传分子标记用于抗病育种，能够极大地促进鸡自身抵抗力的提高，以及减少抗生素等药物在家禽养殖中的使用。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据提供的附图获得其他的附图。
图1附图为TLR1A基因的酶切产物；
图2附图为TLR4基因的酶切产物。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，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，都属于本发明保护的范围。
实施例1SNP位点的筛选和扩增
变异位点的筛选和确定
从NCBI选取鸟类的TLR1A和TLR4基因序列，构建进化树。基于PAML软件包的位点模型(Site Model)分析TLR1A和TLR4各氨基酸位点经受的选择压力，该模型允许每个位点拥有一个ω值。似然率测试(LRT)两个模型，M8和M8a。M8是备择假设(自己估算ω值)，筛选正选择(ω＞1)类位点；而M8a是零假设(ω值固定为1)。若是似然率检验p值显著，则接受备择假设；反之，接受零假设。贝叶斯经验贝叶斯(BEB)可用来计算位点的后验概率，我们将模型比较显著情况下，且BEB＞95％的位点作为正选择候选位点。为了使筛选结果更加可靠，我们还选择了Datamonkey中的FEL、REL、SLAC来检测正选择位点。FEL和REL很好得结合了同义替换率来估算ω值；SLAC通过构建祖先序列并计算整个系统发育树中每个位置的同义和非同义替换数来检测正选择位点。最后将3种及三种以上方法检测到的位点作为正选择位点。
PAML分析结果显示，TLR1A和TLR4各存在一个显著的正选择位点(表1)。这些位点的变异对蛋白质的功能产生影响。根据Ensembl确定碱基突变位置见表2。
表1TLR1A、TLR4基因受正选择情况
表2变异位点情况表
样品采集
用全血平板凝集试验诊断鸡白痢鸡伤寒的方法采集坝上长尾鸡阳性120只，阴性184只；用2.5ml注射器采集翅静脉血2ml，-20℃保存，用于鸡血液基因组DNA的提取(酚-氯仿法)及后续分析。
引物设计及酶切分型
本发明在TLR1A序列的2443位发现T/C变异，在TLR4序列的1147位发现T/C变异，在此位点进行CAPs检测。以鸡血液基因组DNA为模板，根据Ensembl发布的鸡TLR1A、TLR4基因编码区序列，利用primer5.0软件设计错配引物，使其中一条引物3’端和单碱基突变的一种突变型在PCR扩增后形成一个酶切位点，然后用相应的内切酶进行酶切鉴定。限制性内切酶见表1，引物序列、扩增片段见表3。扩增TLR1A片段长度315bp,为TT型。TLR4片段长度220bp，为TT型。TLR1A酶切后可得到2个DNA片段：315bp和290bp(图1)。TLR4酶切后得到2个DNA片段：220bp和196bp(图2)。
表3引物信息表
PCR扩增反应体系，见表4；
表4
| 试剂 | 25μl |
| 2×EsTaqMasterMix(Dye) | 12.5μl |
| ForwardPrimer,10μM | 1μl |
| ReversePrimer,10μM | 1μl |
| TemplateDNA | 1μl |
| ddH2O | 9.5μl |
| 合计 | 25μl |
程序；
TLR1A
TLR4
表5酶切反应体系
| DNA(扩增产物) | 3μl |
| 限制性内切酶 | 0.5μl |
| 10XNEBuffer | 2.5μl |
| ddH2O | 19μl |
| 合计 | 25μl |
运用逻辑回归方法(SNPststs)，在五种不同的遗传模式下(共显性、显性、隐性、超显性和加性)分析单个TLR1A和TLR4基因单个SNP和沙门氏菌易感性的相关性，结果见表6。TLR1A基因结果显示，2443变异位点与沙门氏菌易感性极显著相关(P<0.0001)。在最佳模型(AIC与BIC最小时)—共显性模型下，2443位点基因型T/T在阳性组的频率(9.7％)显著低于对照组的频率(28.9％)，提示T/T型(OR＝0,95％CI＝0.00-NA)可能为沙门氏菌抵抗型。TLR4基因结果显示，1147变异位点与沙门氏菌易感性极显著相关(P<0.0001)。在最佳模型(AIC与BIC最小时)—共显性模型下，1147位点基因型C/C在阳性组的频率(71％)显著高于对照组的频率(58.3％)，提示C/C型(OR＝1)对T/T型(OR＝1)可能为沙门氏菌易感型。
表6TLR1A和TLR4基因多态性位点与鸡沙门氏菌易感性的相关性分析
注：“NA”表示不存在或没有；OR：比值比；CI：可信区间；AIC：赤池信息量准则；BIC贝叶斯信息标准。
实施例2
用鸡白痢鸡伤寒多价染色平板凝集试验抗原检测坝上长尾鸡和太行鸡两个品种各350只，筛选出阳性和阴性个体。用酚-氯仿法抽提DNA，之后根据特异性引物扩增片段并进行基因分型。检测各基因型所占比例如下表7；
表7
本说明书中各个实施例采用递进的方式描述，每个实施例重点说明的都是与其他实施例的不同之处，各个实施例之间相同相似部分互相参见即可。
对所公开的实施例的上述说明，使本领域专业技术人员能够实现或使用本发明。对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的，本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下，在其它实施例中实现。因此，本发明将不会被限制于本文所示的这些实施例，而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。
序列表
<110> 河北科技师范学院
<120> 一种鉴别鸡群对沙门氏菌易感性的SNP位点、筛选及应用
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<170> SIPOSequenceListing 1.0
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Claims (1)
1.一种SNP位点在鉴别对沙门氏菌易感性鸡群中的应用，所述应用为非诊断目的的，其特征在于，所述SNP位点为TLR1A基因第2443位T/C变异和TLR4基因第1147位T/C变异；其中，第2443位T/C变异所在的特征序列为：TATAACCCAA[T/C]GTCTGATTAA，如SEQ ID NO.1所示；第1147位T/C变异所在的特征序列为：GAGTCTAACT[T/C]ACCTGGAGGT，如SEQ ID NO.2所示。
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543227A (zh) * | 2016-01-11 | 2016-05-04 | 山东农业大学 | 一种鸡肠炎沙门氏菌感染抗性性状分子标记gga-miR-1662及其检测方法 |
| CN110004234A (zh) * | 2019-04-19 | 2019-07-12 | 中国农业科学院北京畜牧兽医研究所 | 鸡抗沙门氏菌感染相关的miRNA及其应用 |
| CN110079606A (zh) * | 2019-03-22 | 2019-08-02 | 云南农业大学 | 利用防御素基因标记提高鸡群沙门氏菌抵抗力的方法 |
| WO2021097144A2 (en) * | 2019-11-12 | 2021-05-20 | Actym Therapeutics, Inc. | Immunostimulatory bacteria delivery platforms and their use for delivery of therapeutic products |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543227A (zh) * | 2016-01-11 | 2016-05-04 | 山东农业大学 | 一种鸡肠炎沙门氏菌感染抗性性状分子标记gga-miR-1662及其检测方法 |
| CN110079606A (zh) * | 2019-03-22 | 2019-08-02 | 云南农业大学 | 利用防御素基因标记提高鸡群沙门氏菌抵抗力的方法 |
| CN110004234A (zh) * | 2019-04-19 | 2019-07-12 | 中国农业科学院北京畜牧兽医研究所 | 鸡抗沙门氏菌感染相关的miRNA及其应用 |
| WO2021097144A2 (en) * | 2019-11-12 | 2021-05-20 | Actym Therapeutics, Inc. | Immunostimulatory bacteria delivery platforms and their use for delivery of therapeutic products |
Non-Patent Citations (3)
| Title |
|---|
| Allelic Variation in TLR4 Is Linked to Susceptibility to Salmonella enterica Serovar Typhimurium Infection in Chickens;Gary Leveque等;《INFECTION AND IMMUNITY》;20030531;第71卷(第3期);第1116-1124页 * |
| rs15032440 (SNP);佚名;《Ensembl genome browser 105》;20211231;第1-2页 * |
| 太行鸡脾脏SRBC免疫应答基因的筛选与分析;耿立英;张传生;李祥龙;赵书雨;;中国畜牧杂志;20160110(第01期);第8-14页 * |
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