CAJ | 학술논문

小肠型脂肪酸结合蛋白对长链脂肪酸具有高度的亲和力,参与脂肪酸的吸收和细胞内转运.利用cDNA末端快速扩增(RACE)技术并结合同源克隆策略,克隆到了编码猪小肠型脂肪酸结合蛋白基因(I-FABP)的全长cDNA序列(GenBank接受号:AY960624),并对系统发育关系等进行了生物信息学分析.猪I-FABP基因的cDNA序列全长614 bp,其中包括399 bp的开放式读码框(ORF),43 bp的5'末端非编码区(5'URT)和172 bp的3'末端非编码区(3'URT),编码132个氨基酸残基蛋白,在氨基酸水平上与其他物种的I-FABP具有高度的同源性.以邻接法(Neigbor-Joining,NJ)所构建的系统发育关系表明,猪I-FABP与其他物种的I-FABP属于同一类群,且与人的遗传距离最近.Northern杂交和半定量RT-PCR分析发现,猪I-FABP在猪体组织中出现约620 bp大小的转录本,且在猪体组织中广泛存在,但在小肠组织中表达量最为丰富.
소장형지방산결합단백대장련지방산구유고도적친화력,삼여지방산적흡수화세포내전운.이용cDNA말단쾌속확증(RACE)기술병결합동원극륭책략,극륭도료편마저소장형지방산결합단백기인(I-FABP)적전장cDNA서렬(GenBank접수호:AY960624),병대계통발육관계등진행료생물신식학분석.저I-FABP기인적cDNA서렬전장614 bp,기중포괄399 bp적개방식독마광(ORF),43 bp적5'말단비편마구(5'URT)화172 bp적3'말단비편마구(3'URT),편마132개안기산잔기단백,재안기산수평상여기타물충적I-FABP구유고도적동원성.이린접법(Neigbor-Joining,NJ)소구건적계통발육관계표명,저I-FABP여기타물충적I-FABP속우동일류군,차여인적유전거리최근.Northern잡교화반정량RT-PCR분석발현,저I-FABP재저체조직중출현약620 bp대소적전록본,차재저체조직중엄범존재,단재소장조직중표체량최위봉부.
The intestinal fatty acid-binding protein (I-FABP) shows binding specificity for long-chain fatty acids and is proposed to be involved in the uptake of dietary fatty acids and their intracellular transport. In this study, the full-length cDNA of I-FABP was cloned from pig intestine by homology cloning approach combined with 3' and 5' RACE. Sequence analysis and bioinformatics study showed that this cDNA contained 614 nucleotides, with a 399 bp open reading frame (ORF) flanked by a 43 bp 5' UTR and a172 bp 3' UTR. The encoded 132 amino acids of pig I-FABP with a molecular weight of approximately 15 kDa shared a high sequence identity of 68%-85% with those of other species. In addition, the phylogenetical analysis also indicated that the pig I-FABP was in the same branch with those of other species. The tissue-specific expression of pig I-FABP was measured by Northern hybridization and semi-quantitative RT-PCR. The results demonstrated that pig I-FABP mRNA was extensively present in various tissues, but I-FABP transcript of approximately 620 bp was more abundant in intestine than in other tissues.