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利用逆转录病毒表达载体进行cDNA克隆：pp60c-src cDNA克隆的产生

cDNA cloning with a retrovirus expression vector: generation of a pp60c-src cDNA clone.

作者信息

Kaplan P L, Simon S, Cartwright C A, Eckhart W

出版信息

J Virol. 1987 May;61(5):1731-4. doi: 10.1128/JVI.61.5.1731-1734.1987.

DOI:10.1128/JVI.61.5.1731-1734.1987
PMID:2437323
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC254166/
Abstract

We used a murine retroviral expression vector, containing a genomic clone of the chicken c-src gene, a bacterial origin of replication, and a selectable marker, to remove 10 introns from the c-src gene. All 10 introns were removed accurately, and no mutations were introduced. The processed gene encoded a functional pp60c-src protein tyrosine kinase.

摘要

我们使用了一种鼠逆转录病毒表达载体，其包含鸡c-src基因的基因组克隆、细菌复制起点和一个选择标记，以从c-src基因中去除10个内含子。所有10个内含子均被准确去除，且未引入突变。加工后的基因编码一种功能性的pp60c-src蛋白酪氨酸激酶。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/35b7/254166/4c19662ff669/jvirol00096-0443-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/35b7/254166/0ded9510d429/jvirol00096-0442-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/35b7/254166/4c19662ff669/jvirol00096-0443-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/35b7/254166/0ded9510d429/jvirol00096-0442-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/35b7/254166/4c19662ff669/jvirol00096-0443-a.jpg

相似文献

1
cDNA cloning with a retrovirus expression vector: generation of a pp60c-src cDNA clone.利用逆转录病毒表达载体进行cDNA克隆：pp60c-src cDNA克隆的产生
J Virol. 1987 May;61(5):1731-4. doi: 10.1128/JVI.61.5.1731-1734.1987.
2
Retrovirus shuttle vector for study of kinase activities of pp60c-src synthesized in vitro and overproduced in vivo.用于研究体外合成并在体内过量表达的pp60c-src激酶活性的逆转录病毒穿梭载体。
Mol Cell Biol. 1986 Jun;6(6):2033-40. doi: 10.1128/mcb.6.6.2033-2040.1986.
3
pp60c-src tyrosine kinase, myristylation, and modulatory domains are required for enhanced mitogenic responsiveness to epidermal growth factor seen in cells overexpressing c-src.pp60c-src酪氨酸激酶、肉豆蔻酰化和调节结构域是过表达c-src的细胞中对表皮生长因子增强的促有丝分裂反应性所必需的。
Mol Cell Biol. 1989 Apr;9(4):1536-44. doi: 10.1128/mcb.9.4.1536-1544.1989.
4
The structurally distinct form of pp60c-src detected in neuronal cells is encoded by a unique c-src mRNA.在神经元细胞中检测到的结构不同的pp60c-src形式由一种独特的c-src信使核糖核酸编码。
Mol Cell Biol. 1987 Nov;7(11):4142-5. doi: 10.1128/mcb.7.11.4142-4145.1987.
5
Avian pp60c-src is more active when expressed in yeast than in vertebrate fibroblasts.禽源pp60c-src在酵母中表达时比在脊椎动物成纤维细胞中更具活性。
Oncogene Res. 1987 Sep-Oct;1(4):297-310.
6
Neuronal pp60c-src contains a six-amino acid insertion relative to its non-neuronal counterpart.
Science. 1987 Jul 24;237(4813):411-5. doi: 10.1126/science.2440106.
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Surgery. 1997 Aug;122(2):501-7. doi: 10.1016/s0039-6060(97)90044-1.
8
Structure and expression of STK, a src-related gene in the simple metazoan Hydra attenuata.在简单后生动物细螅中，一种与src相关的基因STK的结构与表达
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Murine c-fms cDNA: cloning, sequence analysis and retroviral expression.
Oncogene Res. 1987 Sep-Oct;1(4):311-24.
10
From c-src to v-src, or the case of the missing C terminus.从c-src到v-src，或者C端缺失的情况。
Cancer Surv. 1986;5(2):159-72.

引用本文的文献

1
Mutation of a cysteine residue in polyomavirus middle T antigen abolishes interactions with protein phosphatase 2A, pp60c-src, and phosphatidylinositol-3 kinase, activation of c-fos expression, and cellular transformation.多瘤病毒中T抗原的一个半胱氨酸残基发生突变，会消除与蛋白磷酸酶2A、pp60c-src和磷脂酰肌醇-3激酶的相互作用，c-fos表达的激活以及细胞转化。
J Virol. 1993 Apr;67(4):1945-52. doi: 10.1128/JVI.67.4.1945-1952.1993.
2
Amino-terminal regions of polyomavirus middle T antigen are required for interactions with protein phosphatase 2A.多瘤病毒中T抗原的氨基末端区域是与蛋白磷酸酶2A相互作用所必需的。
J Virol. 1995 Jun;69(6):3729-36. doi: 10.1128/JVI.69.6.3729-3736.1995.
3

本文引用的文献

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Overexpression of the c-src protein does not induce transformation of NIH 3T3 cells.c-src蛋白的过表达不会诱导NIH 3T3细胞发生转化。
Proc Natl Acad Sci U S A. 1984 Nov;81(22):7071-5. doi: 10.1073/pnas.81.22.7071.
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An unusual 5' splice sequence is efficiently utilized in vivo.一种不寻常的5'剪接序列在体内被有效利用。
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Structure and sequence of the cellular gene homologous to the RSV src gene and the mechanism for generating the transforming virus.
Mutation of a site of tyrosine phosphorylation in the lymphocyte-specific tyrosine protein kinase, p56lck, reveals its oncogenic potential in fibroblasts.
淋巴细胞特异性酪氨酸蛋白激酶p56lck中一个酪氨酸磷酸化位点的突变揭示了其在成纤维细胞中的致癌潜力。
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A retroviral promoter is sufficient to convert proto-src to a transforming gene that is distinct from the src gene of Rous sarcoma virus.逆转录病毒启动子足以将原癌基因src转化为一个与劳氏肉瘤病毒的src基因不同的转化基因。
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6
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J Virol. 1990 Apr;64(4):1734-44. doi: 10.1128/JVI.64.4.1734-1744.1990.
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Analysis of cDNAs of the proto-oncogene c-src: heterogeneity in 5' exons and possible mechanism for the genesis of the 3' end of v-src.原癌基因c-src的cDNA分析：5'外显子的异质性及v-src 3'末端产生的可能机制。
Mol Cell Biol. 1991 Aug;11(8):4165-76. doi: 10.1128/mcb.11.8.4165-4176.1991.
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Functional asymmetry of the regions juxtaposed to the membrane-binding sequence of polyomavirus middle T antigen.与多瘤病毒中T抗原膜结合序列相邻区域的功能不对称性。
Mol Cell Biol. 1992 Nov;12(11):5050-8. doi: 10.1128/mcb.12.11.5050-5058.1992.
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Activated lck tyrosine protein kinase stimulates antigen-independent interleukin-2 production in T cells.活化的lck酪氨酸蛋白激酶刺激T细胞中抗原非依赖性白细胞介素-2的产生。
Mol Cell Biol. 1992 Oct;12(10):4724-32. doi: 10.1128/mcb.12.10.4724-4732.1992.
与劳氏肉瘤病毒src基因同源的细胞基因的结构与序列以及产生转化病毒的机制。
Cell. 1983 Mar;32(3):881-90. doi: 10.1016/0092-8674(83)90073-9.
4
Splicing of intervening sequences introduced into an infectious retroviral vector.引入感染性逆转录病毒载体中的间隔序列的剪接。
J Mol Appl Genet. 1982;1(6):547-59.
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Loss of intervening sequences in genomic mouse alpha-globin DNA inserted in an infectious retrovirus vector.插入感染性逆转录病毒载体的基因组小鼠α-珠蛋白DNA中居间序列的缺失。
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Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.利用处于SV40早期区域启动子控制下的细菌基因将哺乳动物细胞转化为抗生素抗性细胞。
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Molecular cloning and characterization of the chicken gene homologous to the transforming gene of Rous sarcoma virus.与劳氏肉瘤病毒转化基因同源的鸡基因的分子克隆与特性分析
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Monoclonal antibodies to Rous sarcoma virus pp60src react with enzymatically active cellular pp60src of avian and mammalian origin.针对劳氏肉瘤病毒pp60src的单克隆抗体可与源自禽类和哺乳动物的具有酶活性的细胞pp60src发生反应。
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Nucleotide sequence of Moloney murine leukaemia virus.莫洛尼鼠白血病病毒的核苷酸序列。
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High-efficiency gene transfer into mammalian cells: generation of helper-free recombinant retrovirus with broad mammalian host range.高效基因导入哺乳动物细胞：产生具有广泛哺乳动物宿主范围的无辅助病毒重组逆转录病毒。
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