title: Functional analysis of the promoter of the heat shock cognate 70 gene of the Pacific white shrimp, Litopenaeus vannamei.
abstract: Current knowledge on cis-regulatory elements of immune genes of the Pacific white shrimp (Litopenaeus vannamei) is poor. In this study, we identified the promoter of the heat shock cognate protein 70 (HSC70) gene of L. vannamei (lvhsc70). The promoter activity of lvhsc70 promoter was analyzed in insect sf9 cell lines. First, the putative promoter sequence was proved to be able to drive the expression of reporter EGFP gene successfully. Then serial deletion experiments were conducted to investigate functional transcription elements in the promoter region. The results revealed that both positive and negative transcription elements existed in this region. These results are quite different from the previous report on the promoter of HSC70 gene in Penaeus monodon (pmhsc70), where only positive transcription elements were indicated. The sequences that are not conserved between the promoters of lvhsc70 and pmhsc70 might contribute to the differences. Finally, we tested the effect of a putative "NF-κb binding site" in the promoter and, surprisingly, found that deletion of this site would result in a significantly enhancement of the expression of reporter genes, while the underlying mechanisms remain unrevealed. Our results would provide supports for future studies to identify the functional transcription elements in the lvhsc70 promoter and to expand our knowledge on regulation of innate immune genes in penaeid shrimp.
authors.affiliation: Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.
authors.forename: Cui
authors.initials: C
authors.lastname: Zhao
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Zhang
authors.forename: Fuhua
authors.initials: F
authors.lastname: Li
authors.forename: Pin
authors.initials: P
authors.lastname: Huan
authors.forename: Jianhai
authors.initials: J
authors.lastname: Xiang
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017385:Sequence Homology; D061987:Sf9 Cells; D018411:Spodoptera
chemical_list: D060829:Arthropod Proteins; D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger