Premium
Utilization of a Potentially Universal Downstream Primer in the Rapid Identification and Characterization of Vβ Genes from Two New Human Vλ Gene Families
Author(s) -
DEFTOS M.,
SOTOGIL R.,
QUAN M.,
OLEE T.,
CHEN P. P.
Publication year - 1994
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1994.tb03345.x
Subject(s) - primer (cosmetics) , gene , genetics , biology , polymerase chain reaction , cloning (programming) , computational biology , germline , clone (java method) , gene family , genome , chemistry , organic chemistry , computer science , programming language
Polymerase chain reaction (PCR) has increased dramatically the speed of cloning and characterizing numerous genes. However, its application to identifying and analysing new germline Ig‐variable (V) gene families has been hampered by the lack of sequence information in the downstream flanking regions of the concerned V genes, which are deleted during V(D)J rearrangements. To circumvent this problem, the possibility was explored that a degenerate downstream primer may be used in conjunction with a specific upstream primer, to clone members of new Vβ gene families, as much less is known about Vλ genes than Vh and Vk genes in humans. Firstly the feasibility and the specificity of a degenerate primer was examined by comparing it with an established downstream primer in amplifying known Vλ1 genes. The results were positive. Thus, the degenerate primer was used to clone and characterize germline Vλ genes of the recently defined Vλ8 and Vλ9 gene families. This current strategy may help speed up the identification and characterization of all human Vλ genes. Moreover, a similar strategy can be applied to identify and characterize rapidly new V genes of either known or unknown Ig and T‐cell receptor V gene families.