The beta-actin gene promoter of rohu carp (Labeo rohita) drives reporter gene expressions in transgenic rohu and various cell lines, including spermatogonial stem cells
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Hirak Kumar Barman
,
Ramya Mohanta
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.
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Articles in the same Issue
- Biochemical evidence for Ca2+-independent functional activation of hPLSCR1 at low pH
- Catabolism of chondroitin sulfate
- The effect of hypoxia on PGE2-stimulated cAMP generation in HMEC-1
- Is there a connection between inflammation, telomerase activity and the transcriptional status of telomerase reverse transcriptase in renal failure?
- The beta-actin gene promoter of rohu carp (Labeo rohita) drives reporter gene expressions in transgenic rohu and various cell lines, including spermatogonial stem cells
- Molecular machines – a new dimension of biological sciences
- Therapeutic potential of PACAP for neurodegenerative diseases
- A newly isolated yeast as an expression host for recombinant lipase
- Identification of shorter length lamin A protein in mouse ear cartilage tissue
- Relationship among IL-6, LDL cholesterol and lipid peroxidation
- Study strategies for long non-coding RNAs and their roles in regulating gene expression
