CAJ | 학술논문

根据铁蛋白基因的保守序列,搜索GenBank数据库中华鲟的EST数据库得到一条同源序列.通过RT-PCR的方法对该序列进行扩增,修改其测序错误,获得中华鲟铁蛋白亚基cDNA全长,经过注释提交GenBank数据库,获取序列登录号EU348782.该cDNA长度为896bp,包含531bp的完整编码区,推测编码的蛋白质为176aa,分子量为20339.9Mr,理论等电点为5.66.它和大两洋鲑鱼铁蛋白序列同源性最高,达到82.9%.该基闪在中华鲟肝脏、胰脏、肌肉、脑、心脏、鳃和胃粘膜等多种组织表达,在胰脏和心脏中表达量较高,在肌肉组织中表达较低.根据同源模建的方法得到该蛋白质三维结构,其包括5个α螺旋和10个转角结构,和人、蛙和细菌的铁蛋白均能很好的叠合,表现了很高的相似性,表明该蛋白结构和功能在基因进化中的高度保守性.
근거철단백기인적보수서렬,수색GenBank수거고중화심적EST수거고득도일조동원서렬.통과RT-PCR적방법대해서렬진행확증,수개기측서착오,획득중화심철단백아기cDNA전장,경과주석제교GenBank수거고,획취서렬등록호EU348782.해cDNA장도위896bp,포함531bp적완정편마구,추측편마적단백질위176aa,분자량위20339.9Mr,이론등전점위5.66.타화대량양해어철단백서렬동원성최고,체도82.9%.해기섬재중화심간장、이장、기육、뇌、심장、새화위점막등다충조직표체,재이장화심장중표체량교고,재기육조직중표체교저.근거동원모건적방법득도해단백질삼유결구,기포괄5개α라선화10개전각결구,화인、와화세균적철단백균능흔호적첩합,표현료흔고적상사성,표명해단백결구화공능재기인진화중적고도보수성.
According to the conserved sequence of the ferritin gene, a homologous sequence was obtained from the EST database through a BLAST search against the GenBank database. This sequence was amplified with the method of RT-PCR, false sequencing was corrected, and full length cDNA of the ferritin subunit from the Chinese sturgeon was obtained. After being submitted to the GenBank database, the sequence accession number EU348782 was assigned. With the length of 896 bp, this cDNA includes entire coding regions of 531 bp, which encodes 176 amino acids (aa). The molecular weight was predicted to be 20339.9Mr and the theoretical isoelectrie point 5.66. It shares 82.9% protein sequence homology with the ferritin of the Atlantic salmon. This gene is expressed in many organs of the Chinese sturgeon, for example, the liver, pancreas, muscle, brain, heart and gastric mucosa. The highest expression level was found in the pancreas and the heart, while the muscular tissue showed the lowest. Homology modeling was used to predict the 3-D structure of the protein, which included 5 alpha helices and 10 turns. The ferritin protein structure could be overlapped and showed high similarity with that of human, frog and bacteria. It was revealed that this kind of ferritin was highly conserved in structure and function.