Kajimura Y, Toyoda H, Sato M, Miyakoshi S, Kaplan S A, Ike Y, Goyert S M, Silver J, Hawke D, Shively J E
DNA. 1983;2(3):175-82. doi: 10.1089/dna.1983.2.175.
The recent development of the amino acid microsequence technique allows us to obtain partial sequence information using an extremely small amount of protein. Two sets of mixed oligonucleotide probes were chemically synthesized using the amino acid sequence information for the heavy chain of human HLA-DR antigen obtained by the microsequence technique. These two hybridization probes were used to screen cDNA clones constructed from cytoplasmic poly(A)+ mRNA from a human B lymphoblastoid homozygous cell line (LG-2). Of the 10,000 clones screened, two clones hybridized with the probes. DNA sequence analysis showed that the longer one of the two cDNA clones was 1183 nucleotides long, including the entire coding region, the signal peptide region, and the complete 3'-noncoding region. The deduced amino acid sequence of the HLA-DR alpha chain is identical to that of other cell lines with a different HLA-DR typing. However, several nucleotide differences are found in the 3'-untranslated region compared with that of other DR haplotypes.
氨基酸微序列技术的最新进展使我们能够使用极少量的蛋白质获得部分序列信息。利用通过微序列技术获得的人HLA-DR抗原重链的氨基酸序列信息，化学合成了两组混合寡核苷酸探针。这两种杂交探针用于筛选由人B淋巴母细胞纯合细胞系（LG-2）的细胞质聚腺苷酸加（poly(A)+）mRNA构建的cDNA克隆。在筛选的10000个克隆中，有两个克隆与探针杂交。DNA序列分析表明，两个cDNA克隆中较长的一个长1183个核苷酸，包括整个编码区、信号肽区和完整的3'非编码区。HLA-DRα链的推导氨基酸序列与其他具有不同HLA-DR分型的细胞系相同。然而，与其他DR单倍型相比，在3'非翻译区发现了几个核苷酸差异。
