Molecular cloning, characterization and expression analysis of grass carp (Ctenopharyngodon idellus) NF45 (ILF2) cDNA, a subunit of the nuclear factor of activated T-cells (NF-AT).
NF45 (ILF2) and NF90 (ILF3) regulate the IL-2 gene transcription via interaction with the antigen receptor response element. Much work on NF45 has been done in human and mammals while little in fish. In the present study, we have cloned and characterized the full-length cDNA of NF45 in grass carp (Ctenopharyngodon idellus). The grass carp NF45 cDNA of 1563bp contains a short 5'UTR of 24bp, a 3'UTR of 375bp and an open reading frame of 1164bp coding for a protein of 387 aa with a predicted molecular mass of 42.8kDa. The encoded protein shares 86.3-96.7% identities to other homologues. RT-PCR was optimized to estimate the expression level of NF45 in grass carp. The results showed that NF45 is constitutively expressed in most selected tissues, including head kidney, spleen, heart, brain, liver, and gill, although low levels were observed in spleen, liver and gill. The ubiquitous expression of NF45 is consistent with a postulated role in gene regulation at the level of transcription. Stimulating the fish with PHA significantly up-regulated the expression of NF45 in most tissues examined, which potentially indicated that NF45 was involved in the immune responses triggered by PHA.
Lin
Hui-Fang
HF
College of Life Sciences, Zhejiang University, Hangzhou 310012, People's Republic of China.
Shao
Jian-Zhong
JZ
Xiang
Li-Xin
LX
Wang
Hui-Ju
HJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics / Q000502:physiology*; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression / Q000502:physiology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D051903:Nuclear Factor 45 Protein / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D016415:Sequence Alignment / Q000662:veterinary; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D051903:Nuclear Factor 45 Protein