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截短的神经生长因子受体克隆的基因转移导致小鼠成纤维细胞表面表达。

Gene transfer of truncated NGF receptor clones leads to cell surface expression in mouse fibroblasts.

作者信息

Sehgal A, Bothwell M, Chao M

机构信息

Department of Cell Biology and Anatomy, Cornell University Medical College, New York, NY 10021.

出版信息

Nucleic Acids Res. 1989 Jul 25;17(14):5623-32. doi: 10.1093/nar/17.14.5623.

DOI:10.1093/nar/17.14.5623
PMID:2548165
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC318184/
Abstract

Transfection of recombinant bacteriophage clones encoding human NGF receptor sequences resulted in cell surface expression in mouse fibroblasts. Unexpectedly, receptors were expressed even after transfection with phage clones which lack 5' gene sequences. Stable transformants were purified and analyzed in detail. S1 nuclease protection and primer extension analysis revealed that an initiation site lies within an intron sequence in the middle of the receptor gene. A truncated mRNA transcript was detected that allowed for the expression of NGF receptors capable of binding to NGF. Since the original phage clones lacked the first two exons, these results suggest that the normal N-terminal sequences may not be necessary for cell surface expression and binding to NGF.

摘要

编码人神经生长因子（NGF）受体序列的重组噬菌体克隆转染小鼠成纤维细胞后，细胞表面出现了表达。出乎意料的是，即使使用缺乏5'基因序列的噬菌体克隆进行转染，受体仍能表达。对稳定转化体进行了纯化和详细分析。S1核酸酶保护分析和引物延伸分析表明，起始位点位于受体基因中间的一个内含子序列内。检测到一种截短的mRNA转录本，它能够表达能够结合NGF的NGF受体。由于原始噬菌体克隆缺少前两个外显子，这些结果表明正常的N端序列对于细胞表面表达和与NGF结合可能不是必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/4549745dcc5a/nar00131-0209-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/fe1eb02afe9b/nar00131-0205-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/4a2cc724f1e1/nar00131-0205-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/560f54a82e57/nar00131-0206-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/ada4524784bc/nar00131-0207-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/4549745dcc5a/nar00131-0209-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/fe1eb02afe9b/nar00131-0205-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/4a2cc724f1e1/nar00131-0205-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/560f54a82e57/nar00131-0206-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/ada4524784bc/nar00131-0207-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdff/318184/4549745dcc5a/nar00131-0209-a.jpg

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本文引用的文献

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Transforming DNA integrates into the host chromosome.转化DNA整合到宿主染色体中。
Cell. 1981 Jan;23(1):29-39. doi: 10.1016/0092-8674(81)90267-1.
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A single-base change at a splice site in a beta 0-thalassemic gene causes abnormal RNA splicing.β0地中海贫血基因剪接位点的单碱基变化导致异常的RNA剪接。
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