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公开(公告)号：US10113156B2
公开(公告)日：2018-10-30
申请号：US13254223
申请日：2010-03-04
申请人： Alan M. Lambowitz , Sabine Mohr , Georg Mohr , Eman Ghanem
发明人： Alan M. Lambowitz , Sabine Mohr , Georg Mohr , Eman Ghanem
摘要： Stabilized reverse transcriptase fusion proteins including a thermostable reverse transcriptase connected to a stabilizer protein are described. Attaching the stabilizer protein to the thermostable reverse transcriptase stabilizes the fusion protein and can aid in its purification, provide increased solubility, allow for longer storage, or allow the fusion protein to be used under more rigorous conditions such as higher temperature. The stabilized reverse transcriptase fusion protein can also include a linker between the stabilizer protein and the thermostable reverse transcriptase. The stabilized reverse transcriptase fusion proteins are suitable for use in nucleic acid amplification methods such as the reverse transcription polymerase chain reaction and other applications involving cDNA synthesis.
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公开(公告)号：US20120009630A1
公开(公告)日：2012-01-12
申请号：US13254223
申请日：2010-03-04
申请人： Alan M. Lambowitz , Sabine Mohr , Georg Mohr , Eman Ghanem
发明人： Alan M. Lambowitz , Sabine Mohr , Georg Mohr , Eman Ghanem
CPC分类号： C12N9/1276 , C07K2319/00 , C07K2319/24 , C12P19/34 , C12Y207/07049
摘要： Stabilized reverse transcriptase fusion proteins including a thermostable reverse transcriptase connected to a stabilizer protein are described. Attaching the stabilizer protein to the thermostable reverse transcriptase stabilizes the fusion protein and can aid in its purification, provide increased solubility, allow for longer storage, or allow the fusion protein to be used under more rigorous conditions such as higher temperature. The stabilized reverse transcriptase fusion protein can also include a linker between the stabilizer protein and the thermostable reverse transcriptase. The stabilized reverse transcriptase fusion proteins are suitable for use in nucleic acid amplification methods such as the reverse transcription polymerase chain reaction and other applications involving cDNA synthesis.
摘要翻译： 描述了稳定的逆转录酶融合蛋白，包括与稳定蛋白连接的热稳定逆转录酶。 将稳定剂蛋白质连接到热稳定逆转录酶稳定融合蛋白并可以帮助其纯化，提供增加的溶解度，允许更长的储存，或允许融合蛋白在更严格的条件下使用，例如较高的温度。 稳定的逆转录酶融合蛋白还可以包括稳定蛋白和热稳定逆转录酶之间的接头。 稳定的逆转录酶融合蛋白适用于核酸扩增方法，如逆转录聚合酶链反应和涉及cDNA合成的其他应用。
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公开(公告)号：US09012183B2
公开(公告)日：2015-04-21
申请号：US14000513
申请日：2012-02-23
CPC分类号： C12P19/34 , C12N15/1096 , C12Q1/6806 , C12Q2521/107 , C12Q2525/191 , C12Q2525/207
摘要： A method of preparing a DNA copy of a target polynucleotide using template switching is described. The method includes mixing a double stranded template/primer substrate made up of a DNA primer oligonucleotide associated with a complementary oligonucleotide template strand with a target polynucleotide in a reaction medium and adding a suitable amount of a non-retroviral reverse transcriptase to the reaction medium to extend the DNA primer oligonucleotide from its 3′ end to provide a DNA copy polynucleotide. The DNA copy polynucleotide includes a complementary target DNA polynucleotide that is synthesized using the target polynucleotide as a template. Methods of adding nucleotides to the double stranded template/primer substrate are also described. The method can be used to facilitate detection, PCR amplification, cloning, and determination of RNA and DNA sequences.
摘要翻译： 描述了使用模板切换来制备靶多核苷酸的DNA拷贝的方法。 该方法包括将由与互补寡核苷酸模板链相关的DNA引物寡核苷酸组成的双链模板/引物底物与靶多核苷酸混合在反应介质中，并向反应介质中加入适量的非逆转录病毒逆转录酶至 从其3'末端延伸DNA引物寡核苷酸以提供DNA拷贝多核苷酸。 DNA拷贝多核苷酸包括使用靶多核苷酸作为模板合成的互补靶DNA多核苷酸。 还描述了向双链模板/底物底物添加核苷酸的方法。 该方法可用于促进RNA和DNA序列的检测，PCR扩增，克隆和测定。
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公开(公告)号：US20140004569A1
公开(公告)日：2014-01-02
申请号：US14000513
申请日：2012-02-23
IPC分类号： C12P19/34
CPC分类号： C12P19/34 , C12N15/1096 , C12Q1/6806 , C12Q2521/107 , C12Q2525/191 , C12Q2525/207
摘要： A method of preparing a DNA copy of a target polynucleotide using template switching is described. The method includes mixing a double stranded template/primer substrate made up of a DNA primer oligonucleotide associated with a complementary oligonucleotide template strand with a target polynucleotide in a reaction medium and adding a suitable amount of a non-retroviral reverse transcriptase to the reaction medium to extend the DNA primer oligonucleotide from its 3′ end to provide a DNA copy polynucleotide. The DNA copy polynucleotide includes a complementary target DNA polynucleotide that is synthesized using the target polynucleotide as a template. Methods of adding nucleotides to the double stranded template/primer substrate are also described. The method can be used to facilitate detection, PCR amplification, cloning, and determination of RNA and DNA sequences.
摘要翻译： 描述了使用模板切换来制备靶多核苷酸的DNA拷贝的方法。 该方法包括将由与互补寡核苷酸模板链相关的DNA引物寡核苷酸组成的双链模板/引物底物与靶多核苷酸混合在反应介质中，并向反应介质中加入适量的非逆转录病毒逆转录酶至 从其3'末端延伸DNA引物寡核苷酸以提供DNA拷贝多核苷酸。 DNA拷贝多核苷酸包括使用靶多核苷酸作为模板合成的互补靶DNA多核苷酸。 还描述了向双链模板/底物底物添加核苷酸的方法。 该方法可用于促进RNA和DNA序列的检测，PCR扩增，克隆和测定。
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