Construction Of CDNA Library From The Musculature Of Sitchopus Japonicus

Apostichopus japonicus (Selenka) which belongs to Echinodermata, Eleutherozoa, Holothuroidea, spidochirota, Stichopodidae,Apostichopu, is a typical kind of temperate zone species. Apostichopus japonicus possess high nutritional and medical value as they are abundant in proteins and mucopolysaccharide. At present, the researches and reports in the world about Apostichopus japonicus were mainly focused on its medicinal and nutritional value and the technique of adult culture.Howener, researches on its molecular biology and genome structure were rarely reported.In this thesis ,total RNA of muscles of sitchopus japonicus was extracted by TRIZOL Reagent. cDNA library was constructed with SMART cDNA Library Construction Kit. The primary library has a high titer of 3.2×10 6 , in which 96.7% clones are recombinant. The amplified library has a titer of 5.1×109 ,and the length of most cDNA inserted in the library was about 500 - 2500 bp. It concludes that the cDNA library of musculature of sitchopus japonicus has been constructed by using the SMART technique. This cDNA library lays foundation for numbers of expressed sequence tags of sitchopus japonicus and is significant for research on this important aquicultural animal on the level of gene express. It also will help to construct the breeding, growth, resistance, and metabolism rule of sitchopus japonicus.