Analysis Of The Nutritive Components Of The Mandarin Fish And Its Myosin Heavy Chain Gene Cloning And Expression

In recent years, the mandarin fish has become one of the most popular fishspecies in China. In this study, we use the Silver carp as control to analyze thenutritive components of three species of the mandarin fish (S.Kneri, S.Scherzeri andS.Chuatsi) and Silver Carp (Hypophthalmichthys molitrix) with biochemical andproteomics techniques. We then applied RT—PCR/cDNA cloning techniques to clonea myosin heavy chain gene (MHC) from S.Kneri and analyzed its expression profilesat different tissues and developmental stages. The major results are summarized asfollowings:1. Determination of nutrition composition, water: 105℃drying dehydration toanalyze dry weight, The wild S.Kneri is 76.06%, S.Scherzeri is 76.16%, The feedingS.Chuatis is (81.43%) and Silver Carp is 82.10%, Mandarin fish is less than SilverCarp; By using MA ashes Determination of ash content, Mandarin fish is higher thanother fishes; Protein: Trace Kjeldahl method for protein amount determination, theresults revealed that the protein amount of wild S. Kneri and S.Scherzeri are muchhigher than that of S.Chuatis as well as Silver Carp; Fattys: The Suo's extraction of fatcontent, Silver Carp is 5.25%, while S. Chuatis, S.Scherzeri, and S.Kneri rangedfrom 1.09 to 2.18%. Then content of fattys in Mandarin fish is lower than others.fatty acids in fish oil: Determination of fatty acids in fish oil with Gas phasechromatography. In the four species analyzed, they have three major saturated fattyacid(SFA) as C_14:0, C_16:0, C_18:0, and three unsaturated fatty acids, C_18:2, C_20:5, C_22:6.The content of saturated fatty acids in S. Scherzeri, S.Kneri and S. Chuats are 24.4%,29.29% and 35.75% respectively. The components of unsaturated fatty acids are71.96% in S.scherzeris, 67.82% in S.kenri and 54.88% in S.Chuats. However. In theSilver Carp, the amount of SAF is 28.37% and of UFA is 42.05%.2. Cloning the myosin heavy chain (MHC) in full length sequence. The fulllength cDNA of MHC gene in S.Kneri. is 5938bp,3'-race 124bp, two structuredomains, including open reading frame at 5814bp, encoding 1937 amino Acids. Wealso used the techniques of SDS-PAGE/Western blots, as well as RT-PCR amplification to analyze the gene expression profiles at different tissues anddevelopment stages. The MHC gene is expressed specifically in muscle and hearttissues and developmentally from muscle formation stage.