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从斜带石斑Epinepheluscoioides白细胞cDNA文库随机挑取噬菌斑 ,做为模板 ,用λTriplEx5’3’长距离插入子筛选引物 ,进行插入片断的PCR扩增 ,并进行表达序列探针 (ESTs)测定。从中克隆到了血红蛋白β亚基cDNA。cDNA包含长达 4 4 4bp的开放阅读框 ,编码 1 4 7个氨基酸的多肽。用www .expasy .ch网站的computepl/MW程序计算其可能的相对分子质量为 1 65 4 4 ,等电点为 6 .95。其氨基酸序列与草鱼β -珠蛋白III的同源性最高 ,为 73.0 %。在远端和近端的与血红素结合的组氨酸都是保守的。另外 ,与其它鱼类及蛙、鸡和人的 β -珠蛋白比较 ,在血红素结合区及亚基结合区的氨基酸残基均是高度保守的
Plaques were randomly picked from the Epinephelus coioides leukocyte cDNA library of the stained plaques and used as templates to perform PCR amplification of the inserted fragments using λTriplEx 5 ’3’ long-distance insertions, and expressed sequence probes (ESTs) . The hemoglobin β subunit cDNA was cloned from it. The cDNA contains an open reading frame of 44 bp and encodes a polypeptide of 147 amino acids. Using the computepl / MW program at www.expasy.ch, the possible relative molecular mass was calculated to be 1 65 4 4 with an isoelectric point of 6.95. Its amino acid sequence has the highest homology with that of grass carp β - globin III, 73.0%. Both histidine and histidine bound far and proximal are conserved. In addition, the amino acid residues in the heme binding and subunit binding regions are highly conserved compared to other fish and frog, chicken and human beta-globin