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公开(公告)号：US20020090618A1
公开(公告)日：2002-07-11
申请号：US09845157
申请日：2001-05-01
IPC分类号： C12Q001/68 , C12Q001/70 , C12N009/22
CPC分类号： C12N9/1276
摘要： The present invention is in the fields of molecular and cellular biology. The invention is generally related to reverse transcriptase enzymes and methods for the reverse transcription of nucleic acid molecules, especially messenger RNA molecules. Specifically, the invention relates to reverse transcriptase enzymes which have been mutated or modified to increase thermostability, decrease terminal deoxynucleotidyl transferase activity, and/or increase fidelity, and to methods of producing, amplifying or sequencing nucleic acid molecules (particularly cDNA molecules) using these reverse transcriptase enzymes or compositions. The invention also relates to nucleic acid molecules produced by these methods and to the use of such nucleic acid molecules to produce desired polypeptides. The invention also concerns kits comprising such enzymes or compositions.
摘要翻译： 本发明涉及分子和细胞生物学领域。 本发明通常涉及逆转录酶和逆转录核酸分子，特别是信使RNA分子的方法。 具体地，本发明涉及已被突变或修饰以增加热稳定性，降低末端脱氧核苷酸转移酶活性和/或增加保真度的逆转录酶，以及使用这些方法来产生，扩增或测序核酸分子（特别是cDNA分子）的方法 逆转录酶或组合物。 本发明还涉及通过这些方法产生的核酸分子以及这种核酸分子用于产生所需多肽的用途。 本发明还涉及包含这些酶或组合物的试剂盒。
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2.发明申请公开(公告)号：US20030186270A1
公开(公告)日：2003-10-02
申请号：US10292604
申请日：2002-11-13
IPC分类号： C12Q001/68 , C12P019/34 , C12N009/22
CPC分类号： C12N15/1096 , C12N9/1276 , C12N2740/10011 , C12N2740/11011 , C12Q1/6848 , C12Q2527/101 , C12Q2521/301 , C12Q2521/107
摘要： The present invention is generally related to compositions and methods for the reverse transcription of nucleic acid molecules, especially messenger RNA molecules. Specifically, the invention relates to compositions comprising mixtures of polypeptides having reverse transcriptase (RT) activity, and to methods of producing, amplifying or sequencing nucleic acid molecules (particularly cDNA molecules) using these compositions or polypeptides, particularly at temperatures above about 55null C. The invention also relates to nucleic acid molecules produced by these methods, to vectors and host cells comprising these nucleic acid molecules, and to the use of such nucleic acid molecules to produce desired polypeptides. The invention also relates to methods for producing Rous Sarcoma Virus (RSV) and Avian Myeloblastosis Virus (AMV) RTs or other Avian Sarcoma-Leukosis Virus (ASLV) RTs (null and/or null subunits thereof), to isolated nucleic acid molecules encoding such RSV RT, AMV RT or other ASLV RT subunits, to vectors and host cells comprising these isolated nucleic acid molecules and to RSV RT, AMV RT and other ASLV RT subunits produced by these methods. The invention further relates to nucleic acid molecules encoding recombinant heterodimeric RT holoenzymes, particularly heterodimeric RSV RTs, AMV RTs or other ASLV RTs (which may be nullnull RTs, nullnull RTs, or null RTs), vectors (particularly baculovirus vectors) and host cells (particularly insect and yeast cells) comprising these nucleic acid molecules, methods for producing these heterodimeric RTs and heterodimeric RTs produced by these methods. The invention also relates to kits comprising the compositions, polypeptides, or RSV RTs, AMV RTs or other ASLV RTs of the invention.
摘要翻译： 本发明通常涉及核酸分子，特别是信使RNA分子的逆转录的组合物和方法。 具体地，本发明涉及包含具有逆转录酶（RT）活性的多肽的混合物的组合物，以及使用这些组合物或多肽，特别是在高于约55℃的温度下产生，扩增或测序核酸分子（特别是cDNA分子）的方法 本发明还涉及通过这些方法产生的核酸分子，涉及包含这些核酸分子的载体和宿主细胞，以及使用这种核酸分子产生所需多肽。 本发明还涉及用于生产分离的编码这样的核酸分子的Rous肉瘤病毒（RSV）和禽成髓细胞病毒（AMV）RT或其他禽类肉瘤 - 白血病病毒（ASLV）RT（其α和/或β亚基））的方法 RSV RT，AMV RT或其他ASLV RT亚单位，包含这些分离的核酸分子的载体和宿主细胞以及通过这些方法产生的RSV RT，AMV RT和其他ASLV RT亚基。 本发明还涉及编码重组异源二聚体RT全酶，特别是异二聚体RSV RT，AMV RT或其他ASLV RT（其可以是字母表RT，betabeta RT或αRT），载体（特别是杆状病毒载体）和宿主细胞（ 特别是昆虫和酵母细胞），通过这些方法产生的这些异源二聚体RT和异二聚体RT的制备方法。 本发明还涉及包含本发明的组合物，多肽或RSV RT，AMV RT或其他ASLV RT的试剂盒。
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公开(公告)号：US20030198944A1
公开(公告)日：2003-10-23
申请号：US10292662
申请日：2002-11-13
IPC分类号： C12Q001/70 , C12Q001/68 , C12P019/34 , C12N009/22
CPC分类号： C12N15/1096 , C12N9/1276 , C12N2740/10011 , C12N2740/11011 , C12Q1/6848 , C12Q2527/101 , C12Q2521/301 , C12Q2521/107
摘要： The present invention is generally related to compositions and methods for the reverse transcription of nucleic acid molecules, especially messenger RNA molecules. Specifically, the invention relates to compositions comprising mixtures of polypeptides having reverse transcriptase (RT) activity, and to methods of producing, amplifying or sequencing nucleic acid molecules (particularly cDNA molecules) using these compositions or polypeptides, particularly at temperatures above about 55null C. The invention also relates to nucleic acid molecules produced by these methods, to vectors and host cells comprising these nucleic acid molecules, and to the use of such nucleic acid molecules to produce desired polypeptides. The invention also relates to methods for producing Rous Sarcoma Virus (RSV) and Avian Myeloblastosis Virus (AMV) RTs or other Avian Sarcoma-Leukosis Virus (ASLV) RTs (null and/or null subunits thereof), to isolated nucleic acid molecules encoding such RSV RT, AMV RT or other ASLV RT subunits, to vectors and host cells comprising these isolated nucleic acid molecules and to RSV RT, AMV RT and other ASLV RT subunits produced by these methods. The invention further relates to nucleic acid molecules encoding recombinant heterodimeric RT holoenzymes, particularly heterodimeric RSV RTs, AMV RTs or other ASLV RTs (which may be nullnull RTs, nullnull RTs, or null RTs), vectors (particularly baculovirus vectors) and host cells (particularly insect and yeast cells) comprising these nucleic acid molecules, methods for producing these heterodimeric RTs and heterodimeric RTs produced by these methods. The invention also relates to kits comprising the compositions, polypeptides, or RSV RTs, AMV RTs or other ASLV RTs of the invention.
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