线纹海马Hippocampus erectus的tlr21基因结构及其对CpG-ODN的应答特征

doi:10.11978/2018034

Abstract

Abstract:

Toll-like receptors (TLRs) are the major pattern recognition receptors (PRR) of the innate immune system in vertebrates, which can initiate host immune responses by recognizing potential pathogens. The present study focused on the role of Tlr21 involved in immune responses in lined seahorse (Hippocampus erectus). The full-length sequence of the lined seahorse tlr21 gene was cloned. The open reading frame (ORF) of seahorse tlr21 gene includes 2964 base pairs, which is predicted to encode 981 amino acids. The encoding protein had a predicted molecular mass of 246.98 kDa and an estimated isoelectric point of 4.78. Homology comparison and phylogenetic analysis showed that lined seahorse tlr21 shared high sequence identity with its homologs from other species, especially those species in Syngnathidae. Quantitative real-time PCR analysis showed that tlr21 mRNA was widely expressed in various tissues and expressed highest in the kidney. We conducted intraperitoneal CpG-ODNs injections to the lined seahorses. Both CpG-2007 and CpG-HC4040 increased expression levels of tlr21 mRNA in seahorse kidney significantly. In conclusion, these findings indicated that tlr21 initiated the innate immunity in lined seahorse by recognizing CpG DNA.

Key words: tlr21, gene structure, CpG-ODN, immunity, Lined seahorse, tissue expression difference

Yuan ZHANG, Geng QIN, Bo ZHANG, Qiang LIN. The gene structure of tlr21 in lined seahorse, Hippocampus erectus, and its responses to CpG-ODN treatment[J].Journal of Tropical Oceanography, 2019, 38(1): 67-75.

Figures/Tables 6

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References 50

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名称	序列 (5′-3′)	用途
HE tlr21-1F	GCAACCAGCGAAGGGAGA	克隆全长
HE tlr21-1R	GGACACGGAACGGACGAT
HE tlr21-2F	ATCGTCCGTTCCGTGTCC
HE tlr21-2R	CGAGCGCAAGTTGAGGTAG
HE tlr21-3F	CGTCCGTTCCGTGTCCAA
HE tlr21-3R	CAGCGTCACGACGTTTGG
HE tlr21-4F	CCGTTACAACCGCATCCT
HE tlr21-4R	GTCCGTCAGCTCGTCCAA
HE tlr21-5F	CGTCCCTTTATCTCACCAAC
HE tlr21-5R	CAGCATGACCCAGTCTTCG
HE tlr21-6F	CGTCCCTTTATCTCACCAAC
HE tlr21-6R	GTGATGCAGGCACAGTTTC
HE tlr21-7F	GCGTTTCGCTCCTGGTTC
HE tlr21-7R	CCGCCCGAGTTTCTTCAT
HE tlr21-8F	AAATCAAGTACGGCTACTACGC
HE tlr21-8R	TGAGTGTCAGAATCTCCCAAA
tlr13bF	CTTCGCCTGCTTCGGTTAGACAA	荧光定量PCR
tlr13bR	GCTCAAGTTGGACACGGAACGG	荧光定量PCR
β-actinF	TGGGCGTACAACAGGTATCG	内参基因
β-actinR	AGGAGTAGCCACGTTCAGTG	内参基因

CpG-ODN	序列 (5′-3′)
CpG-2000	TCCATGACGTTCCTGCAGTTCCTGACTGG
CpG-2006	TCGTCGTTTTGTCGTTTTGTCGTT
CpG-2007	TCGTCGTTGTCGTTTTGTCGTT
CpG-HC4040	TGACTGTGAACGTTCGAGATGA

The gene structure of tlr21 in lined seahorse, Hippocampus erectus, and its responses to CpG-ODN treatment

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