Torres-Mercado Elineth, Renta Jessicca Y, Rodríguez Yolanda, López-Garriga Juan, Cadilla Carmen L
Department of Chemistry, University of Puerto Rico, Mayagüez, P.O. Box 9019, Mayagüez, Puerto Rico 00681-9019, USA.
J Protein Chem. 2003 Nov;22(7-8):683-90. doi: 10.1023/b:jopc.0000008734.44356.b7.
Hemoglobin II from the clam Lucina pectinata is an oxygen-reactive protein with a unique structural organization in the heme pocket involving residues Gln65 (E7), Tyr30 (B10), Phe44 (CD1), and Phe69 (E11). We employed the reverse transcriptase-polymerase chain reaction (RT-PCR) and methods to synthesize various cDNA(HbII). An initial 300-bp cDNA clone was amplified from total RNA by RT-PCR using degenerate oligonucleotides. Gene-specific primers derived from the HbII-partial cDNA sequence were used to obtain the 5' and 3' ends of the cDNA by RACE. The length of the HbII cDNA, estimated from overlapping clones, was approximately 2114 bases. Northern blot analysis revealed that the mRNA size of HbII agrees with the estimated size using cDNA data. The coding region of the full-length HbII cDNA codes for 151 amino acids. The calculated molecular weight of HbII, including the heme group and acetylated N-terminal residue, is 17,654.07 Da.
来自栉孔扇贝的血红蛋白II是一种对氧有反应的蛋白质，其血红素口袋具有独特的结构组织，涉及Gln65（E7）、Tyr30（B10）、Phe44（CD1）和Phe69（E11）残基。我们采用逆转录聚合酶链反应（RT-PCR）和方法合成了各种cDNA（HbII）。使用简并寡核苷酸通过RT-PCR从总RNA中扩增出一个初始的300bp cDNA克隆。从HbII部分cDNA序列衍生的基因特异性引物用于通过RACE获得cDNA的5'和3'末端。根据重叠克隆估计，HbII cDNA的长度约为2114个碱基。Northern印迹分析表明，HbII的mRNA大小与使用cDNA数据估计的大小一致。全长HbII cDNA的编码区编码151个氨基酸。包括血红素基团和乙酰化N末端残基在内的HbII的计算分子量为17,654.07Da。
