低聚异麦芽糖和酵母细胞壁对中华鳖肝脏中溶菌酶基因表达的半定量分析

目的]探讨低聚异麦芽糖和酵母细胞壁对中华鳖肝脏中溶菌酶基因表达的影响。方法]利用CODEHOP设计简并引物,通过RT-PCR克隆中华鳖溶菌酶基因,然后通过半定量PCR分析低聚异麦芽糖和酵母细胞壁对中华鳖溶菌酶基因在肝脏中表达量的影响。结果]通过克隆中华鳖溶菌酶基因,获得303 bp的溶菌酶基因片段,其GenBank登录号HQ437284,与GenBank登录号Q7LZQ1.3的中华鳖C型溶菌酶氨基酸序列同源性为75%。半定量分析结果表明,低聚异麦芽糖可显著提高中华鳖溶菌酶基因在肝脏中的表达量,而酵母细胞壁对中华鳖溶菌酶基因片段在肝脏中的表达量的影响不显著。结论]该研究可为低聚异麦芽糖和酵母细胞壁在中华鳖养殖中的科学应用以及绿色环保添加剂的开发提供理论依据。

Semi-quantitive Analysis of Isomaltooligosaccharide and Yeast Cell Wall on the Expression of Lysozyme Gene in the Liver of Trionyx sinensis

Objective]The research aimed to study the effects of isomaltooligosaccharide and yeast cell wall on the expression of lysozyme gene in the liver of Trionyx sinensis.Method]A pair of degenerate primers were designed using CODEHOP program.The lysozyme gene in Trionyx sinensis was cloned by using RT-PCR.The effects of isomaltooligosaccharide and yeast cell wall on the expression quantity of lysozyme gene of Trionyx sinensis were studied by using semi-quantitive PCR.Result]A lysozyme gene fragment with the length of 303 bp was obtained by cloning the lysozyme gene in T.sinensis and its accession number on GenBank website was HQ437284.The homology of amino acid sequences between the cloned lysozyme gene fragment and C-type lysozyme of T.sinensis with the accession number on GenBank website of Q7LZQ1.3 was 75%.The semi-quantitive analysis results showed that isomaltooligosaccharide could significantly enhanced the expression quantity of lysozyme gene in the liver of T.sinensis.But yeast cell wall had no significant effect on the expression quantity of lysozyme gene fragment in the liver of T.sinensis.Conclusion]This study could provide theoretical basis for the scientific application of isomaltooligosaccharide and yeast cell wall in the breeding of Trionyx sinensis and the development of green additives.