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利用携带显性选择标记的载体将编码小RNA的爱泼斯坦-巴尔病毒基因转移至人淋巴细胞。

Transfer of the Epstein-Barr virus genes coding for small RNAs to human lymphoid cells with a vector carrying a dominant selectable marker.

作者信息

Rymo L

出版信息

EMBO J. 1983;2(6):839-44. doi: 10.1002/j.1460-2075.1983.tb01511.x.

Abstract

Epstein-Barr virus (EBV)-negative, Burkitt-like lymphoma-derived cells were transformed with a transducing vector (pSV2-gpt) containing the Escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase (XGPRT) and with a derivative of PSV2-gpt that carries the genes for the EBV-associated small RNAs on the EcoRI J fragment of B95-8 EBV DNA inserted at the unique EcoRI site (pJ-gpt). Cells transformed with PSV2-gpt and pJ-gpt express the E. coli gpt gene to approximately the same extent, judged by determinations of the XGPRT activity of cell extracts. Blot hybridisation experiments with restriction endonuclease-cleaved DNA from the transformants have revealed the presence of vector DNA sequences in the cells, at least some of which are most probably integrated into high mol. wt. chromosomal DNA. Northern blot hybridisation analysis of cytoplasmic RNA from pJ-gpt-transformed cells revealed the presence of an EcoRI J DNA complementary RNA species of the same size as the EBV DNA-encoded small RNAs found in EBV-transformed cells.

摘要

用含有编码黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(XGPRT)的大肠杆菌基因的转导载体(pSV2 - gpt)以及PSV2 - gpt的衍生物对爱泼斯坦 - 巴尔病毒(EBV)阴性、伯基特样淋巴瘤衍生细胞进行转化,该衍生物在插入唯一EcoRI位点的B95 - 8 EBV DNA的EcoRI J片段上携带与EBV相关的小RNA基因(pJ - gpt)。通过测定细胞提取物的XGPRT活性判断,用PSV2 - gpt和pJ - gpt转化的细胞表达大肠杆菌gpt基因的程度大致相同。对来自转化体的经限制性内切酶切割的DNA进行印迹杂交实验,结果显示细胞中存在载体DNA序列,其中至少一些很可能整合到高分子量染色体DNA中。对pJ - gpt转化细胞的细胞质RNA进行Northern印迹杂交分析,结果显示存在一种与EBV转化细胞中发现的EBV DNA编码的小RNA大小相同的EcoRI J DNA互补RNA种类。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d34/555198/aa26ebab717d/emboj00259-0039-a.jpg

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