An inducible constitutive expression system in Bombyx mori mediated by phiC31 integrase

Graphical Abstract The present study reports a phiC31 integrase ‐mediated promoter flip system (FLIP) via the attB/attP exchange for the inducible expression of target genes in silkworms. Left inset illustrates the schematic of the FLIP system mediated by the phiC31 integrase‐induced attB/attP exchange. Right inset shows this system activates the efficient a nd inducible expression of DsRed in a silkworm embryo‐derived cell line BmE and silkworm individuals. AbstractInducible gene ‐expression systems play important roles in gene functional assays in the post‐genome era.Streptomyces phage ‐derived phiC31 integrase, which mediates an irreversible site‐specific cassette exchange between the phage attachment site (attP) and the bacterial attachment site (attB), provides a promising option for the construction of a controllable gene ‐expression system. Here, we report a phiC31 integrase‐mediated promoter flip system (FLIP) for the inducible expression of target genes in silkworm (Bombyx mori). First, we constructed a FLIP reporter system, in which a BmAct4 promoter with enhanced translational efficiency was flanked by theattB andattP sites in a head ‐to‐head orientation and further linked in a reverse orientation to a DsRed reporter gene. The coexpression of a C‐terminal modified phiC31‐NLS integrase carrying a simian virus 40 (SV40) nuclear localization signal (NLS) effectively flipped the BmAct4 promoter through anattB/attP exchange, thereby activating ...

Source: Insect Science - September 9, 2020 Category: Biology Authors: Feng Wang, Yan ‐Ting Ji, Chi Tian, Yuan‐Cheng Wang, Shen Xu, Ri‐Yuan Wang, Qian‐Qian Yang, Ping Zhao, Qing‐You Xia Tags: ORIGINAL ARTICLE Source Type: research