Objective To obtain the complete mRNA and promoter sequences of pig FAM213B gene, and provide a basis for studying the mechanism of the FAM213B gene in regulating gestation establishment and embryo development of female pigs.
Method The complete mRNA sequence of the FAM213B gene was obtained using 5′ and 3′RACE methods.The amino acid sequence similarities of different species were analyzed. The gene promoter was cloned, and its transcription activity was detected by the dual luciferase report system in porcine endometrial cells.
Result The mRNA of pig FAM213B gene was 808 bp in full length, including the 5′UTR, CDS and 3′UTR of 67, 609 (including the termination codon) and 132 bp (excluding poly A), respectively. A thioredoxin fold domain was predicted from the 17th to 106th amino acid residues. The obtained transcript and the other two computed transcripts all contained the thioredoxin fold domains, but the tertiary structures of three proteins were highly different. The amino acid sequence of pig FAM213B showed 94.03%, 93.03% and 91.54% similarities with those of goat, cattle and sheep, respectively. The cloned promoter sequence (-2 231/+30) of the FAM213B gene was linked with the dual luciferase report vector, and transfected into the endometrial cells. The promoter fragment could drive the expression of the downstream report gene. There were potential binding sites of transcription factors such as NFκB within the promoter.
Conclusion The complete sequence of the transcript of pig FAM213B gene is 808 bp. The FAM213B protein contains a thioredoxin fold domain. The FAM213B promoter (-2 231/+30) has strong transcription activity in porcine endometrial cells.
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