USE OF FAMILY SPECIFIC LEADER REGION PRIMERS FOR PCR AMPLIFICATION OF THE HUMAN HEAVY-CHAIN VARIABLE REGION GENE REPERTOIRE
USE OF FAMILY SPECIFIC LEADER REGION PRIMERS FOR PCR AMPLIFICATION OF THE HUMAN HEAVY-CHAIN VARIABLE REGION GENE REPERTOIRE
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DOI：
10.1016/0161-5890(92)90100-c
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发表时间：
1992-02-01
影响因子：
3.6
通讯作者：
LEVY, R
中科院分区：
文献类型：
--
作者：
CAMPBELL, MJ;ZELENETZ, AD;LEVY, R
我们设计了一组6个非简并寡核苷酸引物，对应于6个人类V(H)基因家族的每个5'先导区。家族特异性聚合酶链反应扩增的一般策略描述了使用这些引物和一个保守的3'引物对应于框架3，J(H)，或恒定区域。该策略用于分离和测序属于V(H)2和V(H)4家族的新的人类种系V(H)基因。在一定条件下，嵌合V(H)序列是通过“跳跃聚合酶链反应”，结合来自不同种系基因的DNA片段产生的，但这可以通过限制扩增周期的数量来避免。用这些家族特异性引物进行PCR扩增，将有助于研究种系V(H)基因库，以及研究V(H)基因在正常和异常（B细胞恶性肿瘤、自身免疫性疾病等）B细胞群体中的使用。
We have designed a set of six, non-degenerate oligonucleotide primers, corresponding to the 5' leader regions of each of the six human V(H) gene families. A general strategy for family specific polymerase chain reaction amplification is described using these primers and a conserved 3' primer corresponding to frame work 3, J(H), or constant region. This strategy was used to isolate and sequence novel human germline V(H) genes belonging to the V(H)2 and V(H)4 families. Under certain conditions, chimeric V(H) sequences were created by a "jumping polymerase chain reaction", combining DNA segments from different germline genes, but this could be avoided by limiting the number of amplification cycles. PCR amplification with these family specific primers will facilitate studies of the repertoire of germline V(H) genes as well as studies on V(H) gene usage in normal and aberrant (B cell malignancies, autoimmune diseases, etc.) B cell populations.