title: The β-actin gene promoter of rohu carp (Labeo rohita) drives reporter gene expressions in transgenic rohu and various cell lines, including spermatogonial stem cells.
abstract: We previously characterized the β-actin gene promoter of Indian domesticated rohu carp (Labeo rohita) and made a reporter construct via fusion to green fluorescence protein (GFP) cDNA. In this study, the same construct was used to breed transgenic rohu fish. About 20% of the transgenic offspring showed ubiquitous expression of the reporter GFP gene. In a few of the transgenic fish, we documented massive epithelial and/or muscular expression with visible green color under normal light. The expression of GFP mRNA was higher in the muscle tissue of transgenic fish than in that of non-transgenic fish. A highly efficient nucleofection protocol was optimized to transfect proliferating spermatogonial cells of rohu using this reporter construct. The β-actin promoter also drove expressions in HEK293 (derived from human embryonic kidney cells), K562 (human leukemic cells) and SF21 (insect ovarian cells) lines. These findings imply conserved regulatory mechanisms of β-actin gene expression across eukaryotes. Furthermore, the isolated β-actin promoter with consensus regulatory elements has the potential to be used in generating transgenic carp with genes of interest and in basic biology research.
authors.forename: Hirak Kumar
authors.initials: HK
authors.lastname: Barman
authors.forename: Ramya
authors.initials: R
authors.lastname: Mohanta
authors.forename: Swagat Kumar
authors.initials: SK
authors.lastname: Patra
authors.forename: Vemulawada
authors.initials: V
authors.lastname: Chakrapani
authors.forename: Rudra Prasanna
authors.initials: RP
authors.lastname: Panda
authors.forename: Swapnarani
authors.initials: S
authors.lastname: Nayak
authors.forename: Sasmita
authors.initials: S
authors.lastname: Jena
authors.forename: Pallipuram
authors.initials: P
authors.lastname: Jayasankar
authors.forename: Priyanka
authors.initials: P
authors.lastname: Nandanpawar
mesh_terms: D000199:Actins / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D002478:Cells, Cultured; D003530:Cyprinidae / Q000235:genetics*; D029941:Fish Proteins / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D007313:Insecta / Q000235:genetics; D008297:Male; D011401:Promoter Regions, Genetic; D013093:Spermatogonia / Q000166:cytology; D013234:Stem Cells / Q000378:metabolism
chemical_list: D000199:Actins; D029941:Fish Proteins