Construction of ß-actin promoter- rtghI expression vector for brown trout (Salmo trutta macrostigma dumeril, 1858)

Artificial selection techniques have been frequently and traditionally used for improving performance of cultured organisms. The principal of the technique is to select phenotypic characters that show sign of good growth abilities. On the other hand, in genetic manipulation, gene frequency of the desired trait increased in the first generation by altering the genetic structure of the genome of the target species. The aim of this study was to construct a vector that contains the rainbow trout growth hormone gene. For this purpose, the growth hormone gene of Oncorhyncus mykiss and ß-actine promoter of Salmo trutta macrostigma was isolated and after subclonings, the promoter+gene construct were cloned and introduced in to pTRE3G-mCherry eukaryotic expression vector. It was found that ß-actin promoter of Salmo trutta macrostigma contains 1607 bp of the DNA were used for the desired construction. The isolated growth hormone gene of Oncorhyncus mykiss was 4570 bp. For construction of the desired gene cassette, the promoter and the gene DNA sequence (used 4526bp of 4570bp) were put head to tail order next to each other using a computer system. The undesired restriction cutting sites were removed and new restriction sites were introduced for cloning of the gene cassette. The desired whole DNA sequence was synthesized in vitro conditions. After in vitro synthesis of the gene cassette, the promoter+ gene structure were cloned in to pTRE3G-mCherry eukaryotic expression vector. The new vector was named as pMY07. It is expected that the designed structure will improve the growth conditions of the fish.