Multiplex PCR analysis of in vivo‐arising deletion mutations in the hprt gene of human T‐lymphocytes
Multiplex PCR analysis of in vivo‐arising deletion mutations in the hprt gene of human T‐lymphocytes
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人 T 淋巴细胞 hprt 基因体内产生的缺失突变的多重 PCR 分析
DOI：
10.1002/em.2850230203
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发表时间：
1994
影响因子：
2.8
通讯作者：
M. Moore
中科院分区：
文献类型：
--
作者：
J. Fuscoe;L. Zimmerman;K. Harrington‐Brock;M. Moore
采用多重聚合酶链反应（PCR）程序快速有效地评价人T淋巴细胞中次黄嘌呤鸟嘌呤磷酸核糖转移酶（hprt）基因的缺失。使用hprt克隆试验从6名健康男性中分离体内产生的hprt缺陷型T细胞。突变频率范围为9 - 27 × 10−6。从223个突变体的简单的粗细胞提取物进行了分析hprt基因缺失。16例（7.2%）为全基因缺失，22例（9.9%）为部分基因缺失。重排T细胞受体（TCR）-γ基因的单链构象多态性（SSCP）分析显示，单个突变事件的重复分离株导致总基因缺失频率相对较高。基于独特的hprt突变或SSCP-TCR-γ模式，22个部分hprt基因缺失突变体中的18个被确定为独立来源。一半（9/18）的部分缺失突变体仅涉及全部或部分外显子4，这表明hprt基因的该区域容易缺失。影响外显子1（1个突变体）、外显子2（2个突变体）和外显子4（6个突变体）的小缺失无法通过常规Southern印迹分析检测到，可能代表了一种新的、以前未识别的突变类型。这种基因内缺失的现成分离将允许断点连接的表征，并可能提供深入了解DNA断裂和重新连接的重要过程。© 1994 Wiley‐利斯公司
A multiplex polymerase chain reaction (PCR) procedure was adapted for the rapid and efficient evaluation of deletions of the hypoxanthine guanine phosphoribosyltransferase (hprt) gene in human T‐lymphocytes. The hprt clonal assay was used to isolate in vivo‐arising hprt‐deficient T‐cells from six healthy males. Mutant frequencies ranged from 9‐27 × 10−6. Simple crude cellular extracts from 223 mutants were analyzed for hprt gene deletion. Sixteen (7.2%) were found to be due to total gene deletion and 22 (9.9%) were due to partial gene deletion. The relatively high frequency of total gene deletions was caused by replicate isolates of a single mutational event as shown by single‐strand conformation polymorphism (SSCP) analysis of rearranged T‐cell receptor (TCR)‐γ genes. Eighteen of the 22 partial hprt gene deletion mutants were determined to be of independent origin based on a unique hprt mutation or SSCP‐TCR‐γ pattern. One‐half (9/18) of the partial deletion mutants involved all or part of exon 4 alone, suggesting that this region of the hprt gene is prone to deletion. The small deletions effecting exon 1 (1 mutant), exon 2 (2 mutants), and exon 4 (6 mutants) would not have been detected by conventional Southern blot analysis and may represent a new, previously unrecognized class of mutations. The ready isolation of such intragenic deletions will allow the characterization of breakpoint junctions and may provide insights into the important processes of DNA breakage and rejoining. © 1994 Wiley‐Liss, Inc.
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人类体内体细胞突变：测量和分析。
影响因子：
11.1
作者：
Albertini,RJ;Nicklas,JA;O'Neill,JP;Robison,SH
通讯作者：
Robison,SH
对少量二倍体人类细胞裂解物中的 cDNA 进行扩增和直接核苷酸测序。
影响因子：
3.5
作者：
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通讯作者：
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V(D)J 重组酶样活性介导人胎儿 T 淋巴细胞中 hprt 基因缺失。
影响因子：
11.2
作者：
Fuscoe,JC;Zimmerman,LJ;Lippert,MJ;Nicklas,JA;O'Neill,JP;Albertini,RJ
通讯作者：
Albertini,RJ
V(D)J 重组酶介导的成人 T 淋巴细胞中 hprt 基因的缺失。
DOI：
10.1016/0165-7992(92)90116-y
发表时间：
1992
期刊：
Mutation research
影响因子：
--
作者：
Fuscoe,JC;Zimmerman,LJ;Harrington-Brock,K;Burnette,L;Moore,MM;Nicklas,JA;O'Neill,JP;Albertini,RJ
通讯作者：
Albertini,RJ
人 T 淋巴细胞体内 hprt 突变的分子分析。
DOI：
10.1016/0027-5107(89)90178-4
发表时间：
1989
期刊：
Mutation research
影响因子：
--
作者：
Nicklas,JA;Hunter,TC;O'Neill,JP;Albertini,RJ
通讯作者：
Albertini,RJ