An alternative method of promoter assessment by confocal laser scanning microscopy.
A rapid and useful method of promoter activity analysis using techniques of confocal laser scanning microscopy (CLSM) is described in the present study. The activities of some pararetroviral promoters such as CaMV35S (Cauliflower mosaic virus), FMVSgt3 (Figwort mosaic virus sub-genomic transcript) and MMVFLt12 (Mirabilis mosaic virus full-length transcript) coupled to GFP (green fluorescent protein) and GUS (beta-glucuronidase) reporter genes were determined simultaneously by the CLSM technique and other available conventional methods for reporter gene assay based on relevant biochemical and molecular approaches. Consistent and comparable results obtained by CLSM as well as by other conventional assay methods confirm the effectiveness of the CLSM approach for assessment of promoter activity. Hence the CLSM method can be suggested as an alternative way for promoter analysis on the basis of high throughput.
Sahoo
Dipak K
DK
Institute of Life Sciences, Nalco Square, Chandrasekhar Pur, Bhubaneswar, Orissa, India.
Ranjan
Rajiv
R
Kumar
Deepak
D
Kumar
Alok
A
Sahoo
Bhabani S
BS
Raha
Sumita
S
Maiti
Indu B
IB
Dey
Nrisingha
N
D019677:Artificial Gene Fusion; D017796:Caulimovirus / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D011401:Promoter Regions, Genetic
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D005966:Glucuronidase