Parhi Janmejay, Mukherjee S C, Saxena Gopalkrishna, Sahoo Lopamudra, Makesh M
Central Institute of Fisheries Education, Mumbai, India,
Mol Biol Rep. 2014 May;41(5):2979-87. doi: 10.1007/s11033-014-3155-0. Epub 2014 Jan 22.
Genes coding for type-I interferon (I-IFN) has been cloned from Labeo rohita, a commercially important and widely cultured fish in India and South East Asia. In the present study, full-length gene of I-IFN was amplified and sequenced. The sequence analysis revealed that I-IFN consists of 1,786 bp genomic sequence with four introns and five exons and an ORF of 546 bp encoding for a putative protein of 181 amino acids. The mature protein has a molecular weight of 18.97 kDa and consists of 158 amino acids and a signal peptide of 23 amino acids at the N terminus. The sequence carries I-IFN signature motif, one glycosylation site, two conserved cystine amino acids and other conserved amino acids. The sequence showed highest similarity to that of Cyprinus carpio (84%). In silico analysis of the rohu I-IFN protein was done using various bioinformatic tools. The constitutive expression of I-IFN gene was found to be more in spleen compared to gill and kidney in real time PCR assay. Expression of I-IFN increased about 20-fold in cultured kidney cell 2 h after induction with poly I:C and showed maximum expression at 8 h post-induction.
编码I型干扰素（I-IFN）的基因已从印度和东南亚一种具有重要商业价值且广泛养殖的鱼类——印度鲤中克隆出来。在本研究中，扩增并测定了I-IFN的全长基因序列。序列分析表明，I-IFN由1786 bp的基因组序列组成，包含四个内含子和五个外显子，一个546 bp的开放阅读框，编码一个由181个氨基酸组成的假定蛋白。成熟蛋白分子量为18.97 kDa，由158个氨基酸组成，N端有一个23个氨基酸的信号肽。该序列具有I-IFN特征基序、一个糖基化位点、两个保守的半胱氨酸氨基酸和其他保守氨基酸。该序列与鲤鱼（Cyprinus carpio）的序列相似度最高（84%）。使用各种生物信息学工具对印度鲤I-IFN蛋白进行了电子分析。在实时PCR分析中，发现I-IFN基因在脾脏中的组成性表达高于鳃和肾脏。用聚肌胞苷酸（poly I:C）诱导后2小时，培养的肾细胞中I-IFN的表达增加了约20倍，并在诱导后8小时达到最大表达。
