Using SDS-PAGE and ISSR as biochemical markers for assessment the genetic similarity and protein analysis of some cyprinid fish species

Abstract

Genetics similarity and protein analysis of some cyprinid fish were studied using Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Inter-simple sequence repeated (ISSR Markers). Species Pethia nigrofasciatus, Barbonymus schwanenfeldii, Puntius tetrazone and Brachydanio rerio were collected from the fish farms in Damietta to study the genetic variability among them. Eleven ISSR primers were tested to assess the effectiveness of ISSR analysis in discriminating among the four applied fish species. We observed varied size of amplified products depending upon the sequence of ISSR primers and genotypes used. A total of 131 discrete amplified products were obtained (size 79 to 1185 bp approximately) with polymorphism 95%. Out of 131 products, 63 bands were species specific markers indicating high level polymorphism among species. The highest and lowest number of ISSR bands detected for primers ISSR 15 and ISSR16 was 18 and 17 respectively. 7 bands were most relevant as found monomorphic in all four species of family: Cyprinidae. Highest similarity observed between Pethia nigrofasciatus and Barbonymus schwanenfeldii 80% and lowest similarity was between Pethia nigrofasciatus and Brachydanio rerio 31 %. The protein analysis by SDS-PAGE produced 29 bands of molecular weight ranging from 11 to 132 KD with polymorphism 14%. This study concludes that ISSR-based DNA analysis bands and protein profile in muscles from Pethia nigrofasciatus and Barbonymus schwanenfeldii are the most closest species compared molecularly compared with other species used in this study.