CN112899353A - 一种用于dna聚合酶识别特异性的缓冲液及其应用 - Google Patents
一种用于dna聚合酶识别特异性的缓冲液及其应用 Download PDFInfo
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Abstract
本发明提供一种用于DNA聚合酶识别特异性的缓冲液及其应用，所述缓冲溶液包括少于四种碱基类型的dNTPs或少于四种碱基类型的dNTPs的类似物、包含SEQ ID NO：1或SEQ ID NO：2所示的氨基酸序列的DNA聚合酶。通过本发明制备的用于DNA聚合酶识别特异性的缓冲液，可支持含有特定三个或三个以下碱基类型的目标序列的扩增。当目标序列中含有第四个碱基时，扩增反应将终止，因而可明显地控制DNA序列扩增过程产生的非特异性扩增。
Description
技术领域
本发明属于生物技术领域，具体涉及一种用于DNA聚合酶识别特异性的缓冲液及其应用。
背景技术
缓冲溶液是指具有能够维持PH相对稳定性能的溶液。在缓冲溶液中，pH值在一定的范围内不因稀释或外加少量的酸或碱而发生显著的变化，缓冲溶液依据共轭酸碱对及其物质的量不同而具有不同的pH值和缓冲容量。在生化研究工作中，常常需要使用缓冲溶液来维持实验体系的酸碱度。研究工作的溶液体系pH值的变化往往直接影响到DNA聚合酶研究工作的成效。如果DNA聚合酶实验体系的pH值变动或大幅度变动，酶活性就会下降甚至完全丧失，所以配制缓冲溶液是一个不可或缺的关键步骤。尽管DNA聚合酶的缓冲溶液对于DNA聚合酶的活性发挥尤其重要，但是，具体到，如何研发一种特异性识别特定碱基序列的DNA聚合酶的缓冲溶液，能避免DNA聚合酶扩增过程存在的非特异性扩增，在世界范围内还处于一个研究难题。
此外，现有的DNA聚合酶适配的缓冲溶液，在样本中进行聚合酶链式反应时，在样本缓冲溶液中需添加各种dNTPs，才能完成特异性碱基序列识别、扩增或检测。但是对于一些目标序列中含有少于四种碱基的序列的识别过程中，现有的DNA聚合酶普遍存在特异性碱基序列识别的难题，无法有效地扩增出特异性碱基序列。尤为重要的是，现有的DNA聚合酶适配的缓冲溶液，无法完成单一反应体系内超多重PCR的高效和准确的扩增。
因此，亟需开发一种能够用于特异性碱基序列识别的DNA聚合酶的缓冲液。
发明内容
为解决现有技术存在的问题，本发明提供一种用于DNA聚合酶识别特异性的缓冲液、及其制备方法与应用，可支持含有特定三个或三个以下碱基类型的目标序列的扩增。当目标序列中含有第四个碱基时，扩增反应将终止。因此，本发明可明显地控制DNA序列扩增过程产生的非特异性扩增，支持完成单一反应体系内超多重PCR的高效和准确的扩增。
本发明第一方面提供一种用于DNA聚合酶识别特异性的缓冲溶液，其特征在于，所述缓冲溶液包括少于四种碱基类型的dNTPs或少于四种碱基类型的dNTPs的类似物。通过本技术方案，可明显地控制DNA序列扩增过程产生的非特异性扩增。
优选地，如前所述的缓冲溶液，包括DNA聚合酶，所述DNA聚合酶包含SEQ ID NO：1所示的氨基酸序列。通过本技术方案，可明显地控制DNA序列扩增过程产生的非特异性扩增，支持完成单一反应体系内超多重PCR的高效和准确的扩增。
优选地，如前所述的缓冲溶液，包括DNA聚合酶，所述的DNA聚合酶蛋白序列包含在一个或多个位置发生氨基酸取代的SEQ ID NO：1所示的氨基酸序列；其中，所述一个或多个位置选自SEQ ID No:1所示的氨基酸序列中第6位、第8位、第10位、第104位、第205位或其任意组合。通过本技术方案，可明显地控制DNA序列扩增过程产生的非特异性扩增，支持完成单一反应体系内超多重PCR的高效和准确的扩增。当目标序列中含有第四个碱基时，扩增反应将终止。因此，本发明可明显地控制DNA序列扩增过程产生的非特异性扩增。
优选地，如前所述的缓冲溶液包括包括DNA聚合酶，所述的DNA聚合酶蛋白序列包含在多个位置发生氨基酸取代的SEQ ID NO：1所示的氨基酸序列，其中，所述多个位置选自SEQ ID No:1所示的氨基酸序列中第6位、第8位、第10位、第104位和第205位全部发生氨基酸取代。通过本技术方案，可明显地控制DNA序列扩增过程产生的非特异性扩增，支持完成单一反应体系内超多重PCR的高效和准确的扩增。
优选地，在如前所述的缓冲溶液中，所述的SEQ ID No:1所示的氨基酸序列的第6位的氨基酸取代为脯氨酸、甘氨酸、羟脯氨酸、丝氨酸或苏氨酸，和/或第8位的氨基酸取代为异亮氨酸、亮氨酸、缬氨酸、甲硫氨酸、丙氨酸、苯丙氨酸或正亮氨酸取代赖氨酸，和/或第10位的氨基酸取代为苏氨酸或丝氨酸，和/或第104位的氨基酸取代为甘氨酸或脯氨酸，和/或第205位的氨基酸取代为酪氨酸、色氨酸、苯丙氨酸、或苏氨酸。通过本技术方案，可明显地控制DNA序列扩增过程产生的非特异性扩增，支持完成单一反应体系内超多重PCR的高效和准确的扩增。当目标序列中含有第四个碱基时，扩增反应将终止。因此，本发明可明显地控制DNA序列扩增过程产生的非特异性扩增。
优选地，在如前所述的缓冲溶液中，所述的DNA聚合酶包含如SEQ ID No:2所示的氨基酸序列，或由如SEQ IDNo:2所示的氨基酸序列组成。通过本技术方案，可明显地控制DNA序列扩增过程产生的非特异性扩增，支持完成单一反应体系内超多重PCR的高效和准确的扩增。当目标序列中含有第四个碱基时，扩增反应将终止。因此，本发明可明显地控制DNA序列扩增过程产生的非特异性扩增。
优选地，在如前所述的缓冲溶液中，所述DNA聚合酶只识别选自碱基A、T、C和G中的三种碱基构成的核酸序列。
本发明第二方面提供一种识别特异性DNA聚合酶，所述识别特异性DNA聚合酶在应用过程中，采用了权利要求1至6任一权项所述的缓冲溶液。通过本技术方案，可明显地控制DNA序列扩增过程产生的非特异性扩增，支持完成单一反应体系内超多重PCR的高效和准确的扩增。
优选地，在如前所述的识别特异性DNA聚合酶，只识别选自碱基A、T、C和G中的三种碱基构成的核酸序列。通过本技术方案，可明显地控制DNA序列扩增过程产生的非特异性扩增，支持完成单一反应体系内超多重PCR的高效和准确的扩增。当目标序列中含有第四个碱基时，扩增反应将终止。因此，本发明可明显地控制DNA序列扩增过程产生的非特异性扩增。
本发明第三方面提供一种所述的缓冲溶液、或所述识别特异性DNA聚合酶在生物或医学领域的应用。
本发明创造的有益效果：
通过本发明制备的用于DNA聚合酶识别特异性的缓冲液，可支持含有特定三个或三个以下碱基类型的目标序列的扩增。当目标序列中含有第四个碱基时，扩增反应将终止。因此，本发明可明显地控制DNA序列扩增过程产生的非特异性扩增。
附图说明
为了更清楚地说明本发明的技术方案，下面将对实施例中所需要使用的附图作简单地介绍，应当理解，以下附图仅示出了本发明的某些实施例，因此不应被看作是对范围的限定，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，还可以根据这些附图获得其他相关的附图。
图1.识别、扩增仅含有三个碱基的目标片段(Target 1，碱基C不能被DNA聚合酶识别)；
图2.市售的DNA聚合酶扩增仅含有三个碱基的目标片段和正常目标片段；
图3.市售的DNA聚合酶&缓冲液v.s.本实施例的DNA聚合酶&缓冲液的qPCR扩增结果(SYBR Green法)。
具体实施方式
下列实施例中未注明具体条件的实验方法，通常按照国家标准测定。若没有相应的国家标准，则按照通用的国际标准、常规条件、或按照制造厂商所建议的条件进行。
可对本发明提到的特征或实施例提到的特征进行组合。本说明书所揭示的所有特征可与任何组合物形式并用，说明书中所揭示的各个特征，可以任何可提供相同、均等或相似目的的替代性特征取代。因此除有特别说明，所揭示的特征仅为均等或相似特征的一般性例子。
在本发明中，如果没有特别的说明，本文所提到的所有技术特征以及优选特征可以相互组合形成新的技术方案。
为使本发明实现的技术手段、创作特征、达成目的与功效易于明白了解，下面结合具体实施方式，进一步阐述本发明，但本发明包括但不限于这些实施例。
应理解，在本发明范围中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成优选的技术方案。
本文中使用了国际通用的氨基酸的单字母或三字母缩写。
本文所用的术语“多肽”、“肽”和“蛋白”可互换使用，表示多个氨基酸通过肽键连接形成的聚合物。氨基酸可以是天然存在的或人工合成的类似物。
本文中，术语“聚合酶”是包括但不限于如酶命名法所定义的EC2.7.7.7类中的一种酶。
可以根据本领域中已知的程序，如定点诱变或丙氨酸扫描诱变来鉴定多肽中的必需氨基酸。在丙氨酸扫描诱变中，在分子中的每个残基处引入单个丙氨酸突变，并且测试所得突变分子的DNA聚合酶活性以鉴定对分子的活性关键的氨基酸残基。酶或其他生物学相互作用的活性部位还可通过对结构的物理分析来确定，如由下述技术确定：核磁共振、晶体学、电子衍射或光亲和标记，连同对推定的接触位点氨基酸进行突变。还可以从与相关多肽的比对推断鉴别必需氨基酸。
本发明的DNA聚合酶可以在宿主细胞中重组制备并从其分离和纯化。因此，本发明的DNA聚合酶可以被认为是重组酶，特别是分离的重组酶。在某些实施例中，DNA聚合酶是通过重组技术在宿主细胞中制备的，所述宿主细胞不是或不是来自与所述DNA聚合酶来源的生物相同的生物。
本发明的DNA聚合酶可以使用重组DNA技术来产生。可替代地，无细胞表达系统可以用于制备DNA聚合酶。可替代地，可以使用化学合成来产生本发明的DNA聚合酶，从而通过一次一个氨基酸的逐步延伸来产生DNA聚合酶。此类化学合成技术(例如固相合成)在蛋白化学中是众所周知的。
本发明的另一方面提供制备本发明的DNA聚合酶的方法，其包含培养本发明的宿主细胞的步骤。优选的方法包含以下步骤：(i)在适合于表达编码的DNA聚合酶或蛋白的条件下，培养包含一种或多种重组表达载体或一种或多种本发明的核酸分子的宿主细胞；以及任选地(ii)从宿主细胞或从生长培养基/上清液中分离或获得DNA聚合酶或蛋白。此类制备方法还可包含纯化DNA聚合酶或蛋白产物和/或将DNA聚合酶或产物配制成包括至少一种另外的组分例如可接受的缓冲剂或载体的组合物的步骤。
可以使用本领域已知的并且在文献中广泛描述的蛋白的任何纯化技术或其任何组合从宿主细胞/培养基中分开或分离DNA聚合酶。此类技术可以包括例如沉淀，超滤，渗析，各种层析技术，例如，尺寸排阻层析、离子交换层析、亲和层析、电泳、离心分离等。如上所述，可以对本发明的DNA聚合酶进行修饰以携带氨基酸基序或其它蛋白或非蛋白标签，例如聚组氨酸标签(例如His6-标签)，以帮助分离、增溶和/或纯化或鉴定。
另一方面，本发明提供使用本发明的DNA聚合酶扩增核酸(DNA)的方法。通常，所述方法包含提供反应混合物，其包含本发明的DNA聚合酶、模板核酸分子、能够退火至模板核酸分子的一部分的寡核苷酸引物(例如2个或更多个引物，例如2、3、4、5或6个引物)和核苷酸(例如三磷酸脱氧核糖核苷，dNTP)；并在寡核苷酸引物退火至模板核酸分子和所述DNA聚合酶通过聚合一个或多个核苷酸以产生多核苷酸来延伸所述寡核苷酸引物的条件下温育所述反应混合物。合适的条件是本领域众所周知的。核酸扩增的优选的方法是等温扩增方法。本发明的等温扩增方法在恒定温度下进行，并且优选的温度在本文其它地方列出。可选地，检测多核苷酸产物的产生(例如，通过凝胶电泳)。
示例性的等温扩增方法包括环介导等温扩增(LAMP)、滚环扩增(RCA)，链置换扩增(SDA)、多置换扩增(MDA)和交叉引物扩增(CPA)。
对于本发明的其它DNA聚合酶，例如那些基于来自嗜热生物体的序列的聚合酶，恒定温度可以是中等温度至高温，例如选自25℃至65℃，优选40℃至65℃的范围内。
当在反应过程中没有采取任何主动步骤来改变温度时，例如没有热循环时，可以认为温度是恒定的。在此方法期间，“恒”温可能仍会导致例如至多约为5℃，通常不超过3℃或2℃的温度波动。
本发明的DNA聚合酶可以用于即时护理分子诊断平台。
本发明的DNA聚合酶可用于全基因组扩增。
本发明的DNA聚合酶可用于下一代测序方法中。所谓的“下一代”或“第二代”测序方法(参考作为“第一代”方法的桑格(Sanger)双脱氧核苷酸方法)已经普及。这些较新的技术的特征在于产出量高，例如由于使用并行，例如大规模并行测序反应，或通过耗时较少的步骤。各种高产出量测序方法可提供单分子测序，并采用诸如焦磷酸测序、可逆终止子测序、通过结扎的可切割的探针测序、通过结扎的不可切割的探针测序、DNA纳米球和实时单分子测序的技术。
本文中对本发明的DNA聚合酶的引用包括活性片段，除非上下文另有明确说明。
本发明的用途和方法通常在体外进行。
本发明还提供包含本发明的DNA聚合酶的组合物。此类组合物优选包含缓冲剂。任选地，本发明的组合物进一步包含用于进行核酸扩增反应(例如等温扩增反应)的一种或多种必要的试剂，例如能够退火至待扩增的模板DNA的区域的寡核苷酸引物和/或核苷酸(例如dNTP)。通常，组合物将是水性的，并用标准缓冲剂(例如Tris、HEPES等)缓冲。
本发明进一步包括试剂盒，其包含一种或多种本发明的DNA聚合酶、或一种或多种本发明的组合物、或一种或多种本发明的核酸分子、或一种或多种本发明的表达载体、或一种或多种本发明的宿主细胞或病毒。优选地，所述试剂盒用于本文所述的方法和用途，例如，用于核酸扩增方法，例如等温扩增反应。优选地，所述试剂盒包含用于使用试剂盒组分，例如用于核酸扩增的说明书。
本文公开的氨基酸序列及其序列标识符(SEQ ID NO)。
实施例1.一种用于DNA聚合酶识别特异性的缓冲溶液
本发明提供一种用于DNA聚合酶识别特异性的缓冲溶液，将所述缓冲溶液的组分和配比如表1所示进行配置，在25～40℃温度条件下进行混合，制备得到所述DNA聚合酶识别特异性的缓冲溶液。其中，在所述缓冲溶液中，dNTPs只包括三磷酸胸腺嘧啶脱氧核苷酸(dTTP)、三磷酸胞嘧啶脱氧核苷酸(dCTP)和三磷酸鸟嘌呤脱氧核苷酸(dGTP)。
表1.PCR检测体系，总体积50μL
| 组分 | 浓度 | 体积(μL) |
| 人全血 | - | 10 |
| DNA聚合酶 | 20ng/μL | 1.0,2.0,3.0 |
| dNTPs | 10mM each | 1.0 |
| Primer-L | 10μM | 2.0 |
| Primer-R | 10μM | 2.0 |
| Tris-HCl(pH 8.4) | 2M | 0.5 |
| MgCl<sub>2</sub> | 25mM | 4.0 |
| KCl | 1M | 1.0 |
| Triton X-100 | 1％ | 5.0 |
| water | / | 21.5,22.5,23.5 |
在本实施例中，采用的DNA聚合酶蛋白序列为SEQ ID NO：1所示的氨基酸序列。
实施例2.一种用于DNA聚合酶识别特异性的缓冲溶液
本发明提供一种用于DNA聚合酶识别特异性的缓冲溶液，与实施例1不同之处在于，在所述缓冲溶液中，dNTPs只包括三磷酸腺嘌呤脱氧核苷酸(dATP)、三磷酸胞嘧啶脱氧核苷酸(dCTP)和三磷酸鸟嘌呤脱氧核苷酸(dGTP)。
实施例3.一种用于DNA聚合酶识别特异性的缓冲溶液
本发明提供一种用于DNA聚合酶识别特异性的缓冲溶液，与实施例1不同之处在于，在所述缓冲溶液中，dNTPs只包括三磷酸腺嘌呤脱氧核苷酸(dATP)、三磷酸胸腺嘧啶脱氧核苷酸(dTTP)和三磷酸鸟嘌呤脱氧核苷酸(dGTP)。
实施例4.一种用于DNA聚合酶识别特异性的缓冲溶液
本发明提供一种用于DNA聚合酶识别特异性的缓冲溶液，与实施例1不同之处在于，在所述缓冲溶液中，dNTPs只包括三磷酸腺嘌呤脱氧核苷酸(dATP)、三磷酸胸腺嘧啶脱氧核苷酸(dTTP)和三磷酸胞嘧啶脱氧核苷酸(dCTP)。
实施例5：DNA聚合酶酶活性与扩增效率测试
采用实施例1所述的缓冲溶液、和市售的缓冲溶液(采购自天津百伦斯生物技术有限公司，货号bls)，在本实施例中测试了实施例1所述的DNA聚合酶的活性；同时和市售TaqDNA聚合酶(采购自北京索莱宝科技有限公司，货号M1665S)的对目标产物的扩增效率，使用的模板为人工合成的DNA目标片段，扩增产物大小分别为246bp。
扩增产物为246bp体系的引物序列：
S-F：CCCTGGGCTCTGTAAAGAATAGCA(SEQ ID NO：3)；
S-R：ATCAGAGCTTAAACTGGGAAGCTA(SEQ ID NO：4)；
PCR反应体系为：
扩增产物为246bp体系的PCR热循环条件为：95℃，30s；
30cycles*(95℃，15s；58℃，15s；72℃，30s)；72℃，5min。
琼脂糖凝胶电泳结果如图1所示，实验结果通过比对扩增产物条带亮度来体现。
采用实施例1制备的DNA聚合酶，对人工合成的DNA目标片段(SEQ ID NO：5所示的DNA序列和SEQ ID NO：6所示的DNA序列)进行扩增，片段1(Target 1，SEQ ID NO：7所示的DNA序列)为仅含有A、T、G三种碱基的片段；片段2(Target 2，SEQ ID NO：8所示的DNA序列)中含有A、T、C、G四种碱基；NC为negative control。得到的实验结果如图1所示，表明实施例1制备的DNA聚合酶可特异性识别仅含有A、T、G三种碱基的片段1(Target 1)，而不会去识别含有A、T、C、G四种碱基的片段2(Target 2)，具有序列识别的特异性。
Test1的序列：
GGGACCCGAGACATTTCTTATCGTATTGAATGATGGATGAATATATGAATGATGGATGAAATGATGATGATGATGATGATGATGATGATGAATGATGATGAATGAATGATGTTGATGAAATATGTGATGATGATGTAGGTGATGATGATGATGGATGATGAAGGTGATGGATGATGATGATGATGATGATGATGATGATGATGATGATAGATGATGATGATGTAGTCTCGAATTTGACCCTTCGAT(Test1的序列，SEQ ID NO：5)
Test2的序列
GGGACCCGAGACATTTCTTATCGTATCTGAATGATGCGATGAATATATGAATGATGGATGAAATGATGATCGATGATGATGATGATGATGACTGAATGATGATGAATGAATGATGTTGCATGAAATATGCTGATGATGATCGTAGGTGATGATGATGATGGCATGATGAAGGTGATGCGATGATGATGATGATGATGATCGATGATGATGATGACTGATAGATGATGATGATGTAGTCTCGAATTTGACCCTTCGAT(Test2的序列，SEQ ID NO：6)。
采用市售的DNA聚合酶，对人工合成的DNA目标片段(SEQ ID NO：5所示的DNA序列和SEQ ID NO：6所示的DNA序列)进行扩增，片段1(Target1，SEQ ID NO：7所示的DNA序列)为仅含有A、T、G三种碱基的片段；片段2(Target 2，SEQ ID NO：8所示的DNA序列)中含有A、T、C、G四种碱基；NC为negative control。得到的实验结果如图2所示，表明市售的的DNA聚合酶可识别仅含有A、T、G三种碱基的片段1(Target 1)，同时也识别含有A、T、C、G四种碱基的片段2(Target 2)，不具有序列识别的特异性。
ATTGAATGATGGATGAATATATGAATGATGGATGAAATGATGATGATGATGATGATGATGATGATGAATGATGATGAATGAATGATGTTGATGAAATATGTGATGATGATGTAGGTGATGATGATGATGGATGATGAAGGTGATGGATGATGATGATGATGATGATGATGATGATGATGATGATAGATGATGATGATG(target1序列，SEQ ID NO： 7所示序列)；
ATCTGAATGATGCGATGAATATATGAATGATGGATGAAATGATGATCGATGATGATGATGATGATGACTGAATGATGATGAATGAATGATGTTGCATGAAATATGCTGATGATGATCGTAGGTGATGATGATGATGGCATGATGAAGGTGATGCGATGATGATGATGATGATGATCGATGATGATGATGACTGATAGATGATGATGATG(target2序列，SEQ ID NO：8所示序列)。
结果表明，使用实施例1所述的DNA聚合酶缓冲溶液，能成功扩增出含有三个碱基的目标片段，无非特异性扩增。而市售的DNA聚合酶和缓冲溶液，存在非特异性扩增现象。
在本发明的一些实施例中，采用实施例2-4制备的缓冲溶液重复实施例5的实验，同样未出现非特异性扩增的现象。进一步地，采用其它公司购买的缓冲溶液，重复实施例5的实验，出现了非特异性扩增。
在本发明的一些实施例中，发明人在实施例2-4制备的缓冲溶液中的dNTPs的基础上，通过现有技术制备dNTPs的衍生物(曲晓欢，冯旭利.几种经典碱基类似物及应用.重庆大学药学院)，再来重复实施例5的实验，同样未出现非特异性扩增的现象。进一步地，采用其它公司购买的缓冲溶液，重复实施例5的实验，出现了非特异性扩增。
实施例6：多重PCR引物对数测试
采用实施例1所述的缓冲溶液、和市售的缓冲溶液(采购自天津百伦斯生物技术有限公司，货号bls)，在本实施例中测试了实施例1所述的DNA聚合酶的活性；同时和市售TaqDNA聚合酶(采购自北京索莱宝科技有限公司，货号M1665S)进行空白模板条件下的qPCR扩增。设计引物(引物总浓度5μM)变量的对数为24对(采购自ThermoFisher公司，货号：4476135)，分别使用实施例1中所述的缓冲溶液和DNA聚合酶、市售DNA聚合酶和缓冲溶液进行扩增，其他反应条件相同。PCR反应体系如下：特异引物混合物2μL(5μM)；DNA模板/质控品50ng；2×DNA聚合酶工作液(DNA聚合酶用量为0.5U)12.5μL；无核酸酶水将体积补足至25μL。
PCR反应条件：
(1)98℃，激活5min；循环1次；
(2)98℃，变性15s；62℃，退火30s-10min；72℃，延伸2min；(循环30次)；
(3)72℃，延伸10min；循环1次。
注：退火时间随着引物对数增加而增加
如图3所示，当引物对数为24对时，市售DNA聚合酶在30个循环时，已经出现非常明显的非特异性扩增；而实施例1的DNA聚合酶和缓冲液在在100个循环时，仍未出现任何非特异性扩增。
进一步地，发明人对于本实施例1的DNA聚合酶进行优化，采用的DNA聚合酶序列为包含在一个或多个位置发生氨基酸取代的SEQ ID NO：1所示的氨基酸序列，其中所述一个或多个位置选自SEQ ID No:2所示的氨基酸序列中第6位、第8位、第10位、第104位、第205位或其任意组合；进行实施例5或实例例6的实验时，未出现非特异性扩增的现象。进一步地，采用其它公司购买的缓冲溶液，重复实施例5或实例例6的实验时，出现了非特异性扩增。
进一步地，发明人对于本实施例1的DNA聚合酶进行优化，采用的DNA聚合酶序列为SEQ ID No:1所示的氨基酸序列的第6位的氨基酸取代为脯氨酸、甘氨酸、羟脯氨酸、丝氨酸或苏氨酸，和/或第8位的氨基酸取代为异亮氨酸、亮氨酸、缬氨酸、甲硫氨酸、丙氨酸、苯丙氨酸或正亮氨酸取代赖氨酸，和/或第10位的氨基酸取代为苏氨酸或丝氨酸，和/或第104位的氨基酸取代为甘氨酸或脯氨酸，和/或第205位的氨基酸取代为酪氨酸、色氨酸、苯丙氨酸、或苏氨酸；；进行实施例5或实例例6的实验时，未出现非特异性扩增的现象。进一步地，采用其它公司购买的缓冲溶液，重复实施例5或实例例6的实验时，出现了非特异性扩增。
进一步地，发明人对于本实施例1的DNA聚合酶进行优化，所述的DNA聚合酶包含如SEQ ID No:2所示的氨基酸序列或由如SEQ ID No:2所示的氨基酸序列组成；进行实施例5或实例例6的实验时，未出现非特异性扩增的现象。进一步地，采用其它公司购买的缓冲溶液，重复实施例5或实例例6的实验时，出现了非特异性扩增。
以上显示和描述了本发明的基本原理和主要特征和本发明的优点。本行业的技术人员应该了解，本发明不受上述实施例的限制，上述实施例和说明书中描述的只是说明本发明的原理，在不脱离本发明精神和范围的前提下，本发明还会有各种变化和改进，这些变化和改进都落入要求保护的本发明范围内。本发明要求保护范围由所附的权利要求书及其等效物界定。
序列表
<110> 南京普济生物有限公司
<120> 一种用于DNA聚合酶识别特异性的缓冲液及其应用
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Claims (10)
1.一种用于DNA聚合酶识别特异性的缓冲溶液，其特征在于，所述缓冲溶液包括少于四种碱基类型的dNTPs或少于四种碱基类型的dNTPs的类似物。
2.根据权利要求1所述的缓冲溶液，其特征在于，所述的缓冲溶液包括DNA聚合酶，所述DNA聚合酶包含SEQ ID NO：1所示的氨基酸序列。
3.根据权利要求1所述的缓冲溶液，其特征在于，所述的缓冲溶液包括DNA聚合酶，所述的DNA聚合酶蛋白序列包含在一个或多个位置发生氨基酸取代的SEQ ID NO：1所示的氨基酸序列 ；其中，所述一个或多个位置选自SEQ ID No:1所示的氨基酸序列中第6位、第8位、第10位、第104位、第205位或其任意组合。
4.根据权利要求1所述的缓冲溶液，其特征在于，所述的缓冲溶液包括包括DNA聚合酶，所述的DNA聚合酶蛋白序列包含在多个位置发生氨基酸取代的SEQ ID NO：1所示的氨基酸序列 ，其中，所述多个位置选自SEQ ID No:1所示的氨基酸序列中第6位、第8位、第10位、第104位和第205位全部发生氨基酸取代。
5.根据权利要求4所述的缓冲溶液，其特征在于，SEQ ID No:1所示的氨基酸序列的第6位的氨基酸取代为脯氨酸、甘氨酸、羟脯氨酸、丝氨酸或苏氨酸，和/或第8位的氨基酸取代为异亮氨酸、亮氨酸、缬氨酸、甲硫氨酸、丙氨酸、苯丙氨酸或正亮氨酸取代赖氨酸，和/或第10位的氨基酸取代为苏氨酸或丝氨酸，和/或第104位的氨基酸取代为甘氨酸或脯氨酸，和/或第205位的氨基酸取代为酪氨酸、色氨酸、苯丙氨酸、或苏氨酸。
6.根据权利要求4所述的缓冲溶液，其特征在于，所述的DNA聚合酶包含如SEQ ID No:2所示的氨基酸序列，或由如SEQ ID No:2所示的氨基酸序列组成。
7.根据权利要求2至6任一权项所述的缓冲溶液，其特征在于，所述DNA聚合酶只识别选自碱基A、T、C和G中的三种碱基构成的核酸序列。
8.一种识别特异性DNA聚合酶，其特征在于，所述识别特异性DNA聚合酶在应用过程中，采用了权利要求1至6任一权项所述的缓冲溶液。
9.根据权利要求8所述识别特异性DNA聚合酶，其特征在于，所述DNA聚合酶只识别选自碱基A、T、C和G中的三种碱基构成的核酸序列。
10.一种权利要求1至6任一权项所述的缓冲溶液、或权利要求8或9所述识别特异性DNA聚合酶在生物或医学领域的应用。
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2003089637A1 (en) * | 2002-04-17 | 2003-10-30 | The University Of Newcastle_Upon Tyne | Mutation of dna polymerases from archaeobacteria |
| WO2006030174A1 (en) * | 2004-09-15 | 2006-03-23 | The University Of Newcastle | Dna polymerase |
| CN111454926A (zh) * | 2020-05-11 | 2020-07-28 | 南京君华基因科技有限公司 | 一种优化的扩增目标核酸的聚合酶、复合体系及应用 |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003089637A1 (en) * | 2002-04-17 | 2003-10-30 | The University Of Newcastle_Upon Tyne | Mutation of dna polymerases from archaeobacteria |
| WO2006030174A1 (en) * | 2004-09-15 | 2006-03-23 | The University Of Newcastle | Dna polymerase |
| CN111454926A (zh) * | 2020-05-11 | 2020-07-28 | 南京君华基因科技有限公司 | 一种优化的扩增目标核酸的聚合酶、复合体系及应用 |
Non-Patent Citations (1)
| Title |
|---|
| 佚名: "Accession No. WP_011011325.1", 《GENBANK》 * |
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