Schechter I
Proc Natl Acad Sci U S A. 1975 Jul;72(7):2511-4. doi: 10.1073/pnas.72.7.2511.
The mRNA coding for a kappa-type immunoglobulin light (L)-chain and its complementary DNA (cDNA) hybridize with a Crt1/2 of 2.6 x 10(-4) moles of ribonucleotide x liter-1 x sec, forming well-matched duplexes (melting temperature Tm equals 89 degrees). The molecular weight of the cDNA is about 280,000 (840 nucleotides) as determined by alkaline sucrose gradient centrifugation and from the extent of protection of the mRNA by the cDNA from ribonuclease digestion. The cDNA anneals with kappa-type mRNAs of the same and different subgroups with comparable Crt1/2 values, but not with a lambda-type mRNA. Thus, one kappa-type cDNA can be used to quantify the mRNAs coding for all kappa-type L-chains. The values of cDNA hybridized at saturation with various kappa-type mRNAs indicate that: (1) the cDNA is complementary to the entire constant region and to about half of the variable (V)-region; (2) V-regions of similar amino-acid sequence are coded by a similar nucleotide sequence; (3) the nucleic acid probe to one V-region may anneal and quantify V-region genes of members of the same subgroup.
编码κ型免疫球蛋白轻(L)链的信使核糖核酸(mRNA)及其互补脱氧核糖核酸(cDNA)以2.6×10⁻⁴摩尔核糖核苷酸×升⁻¹×秒的Crt1/2进行杂交,形成匹配良好的双链体(解链温度Tm等于89摄氏度)。通过碱性蔗糖梯度离心以及根据cDNA对mRNA免受核糖核酸酶消化的保护程度测定,cDNA的分子量约为280,000(840个核苷酸)。该cDNA与相同和不同亚组的κ型mRNA以相当的Crt1/2值退火,但不与λ型mRNA退火。因此,一种κ型cDNA可用于定量编码所有κ型L链的mRNA。与各种κ型mRNA饱和杂交时的cDNA值表明:(1)该cDNA与整个恒定区以及约一半的可变(V)区互补;(2)氨基酸序列相似的V区由相似的核苷酸序列编码;(3)针对一个V区的核酸探针可能会与同一亚组成员的V区基因退火并进行定量。