GFP EXPRESSION IN PANCREAS OF DEVELOPING FISH EMBRYO UNDER CONTROL OF CARBOXYPEPTIDASE A PROMOTER

Anand Shanker Srivastava National Research Institute of Aquculture Nansei, Mie-516 0193, Japan.

Obtaining the promoter of pancreatic enzyme which works in tissue specific manner will be useful in various fields; e.g. introduction of exogenous genes into pancreas, analysis of organogenesis of pancreas, analysis of circadian rhythm of pancreatic enzyme synthesis e.t.c. Here we report about the cloning of promoter region of carboxypeptidase A (CbA) from flounder, Paralichthys olivaceus (teleostei) and expression analysis of obtained promoter using green fluorescent protein (GFP) vector system. Upstream region of 1800 bp from methionine initiation codan of CbA was PCR amplified from flounder genome using Gene Walker Kit (Clontech). A partial fragment of 1100 bp was ligated with pd2EGFP-1 vector (Clontech). This CbA promoter-GFP vector was microinjected into the fertilized eggs of ice goby, Leucopsarion petersii (teleostei). After seven days of injection, as soon as pancreas differentiated from gut, it started emitting GFP light. Any other organ did not express GFP, indicating that this promoter region of 1100 bp is enough to work in the pancreas-specific gene expression.