共表达3种纤维素降解酶的转基因小鼠模型的建立与分析  

Generation of a Transgenic Mouse Model Co-expressing Three Fibrolytic Enzyme Genes

作　　者：何祖勇[1] 刘志国[1,2] 孙燕霞[1] 丛佩清[1] 陈瑶生[1] 

机构地区：[1]中山大学生命科学学院,有害生物控制与资源利用国家重点实验室,广州510006 [2]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《畜牧兽医学报》2016年第6期1162-1169,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：广东省自然科学基金项目(U1201213);国家转基因动物新品种培育重大专项(2014ZX08006005-005)

摘　　要：本研究旨在应用转基因技术制作能特异性分解纤维素和木质素的转基因单胃动物,对于培育高饲料转化率的家畜具有重要的研究意义。利用2A肽策略构建由EF1α启动子驱动的可同时表达3种能够降解纤维素和木质素的酶(EGXA、BGL4和XYNB)的真核表达载体,并通过原核显微注射技术获得同时表达3种酶的转基因小鼠。经检测发现,2A肽能够介导3种酶同时在HEK293A细胞中正常表达。通过酶活分析、Western blot和免疫组织化学等检测发现,3种酶均能在转基因小鼠消化系统各组织中表达,并且具有正常活性。本研究结果表明,在EF1α启动子的控制下,2A肽能够在小鼠中介导EGXA、BGL4和XYNB的正常表达,并成功建立表达外源纤维素酶和木质素酶的转基因小鼠模型,为培育具有高饲料转化率的动物奠定研究基础。By expressing fibrolytic enzymes targeting cellulose and xylan through transgenic technology,new breeds of monogastric animals with enhanced feed conversion rates could be developed.Here we described the co-expression of cellulase genes EGXA and BGL4,and the xylanase gene XYNB,under the control of EF1α promoter and linked via 2A peptides,in both HEK293 A cells and transgenic mice.Recombinant enzyme expression was detected in gastrointestinal tract as determined by both Western blot and immunohistochemistry,and were functionally active in cellulolytic enzyme assays.Notably,all 3fibrolytic enzymes were highly expressed in the intestine tissue of transgenic mice.In summary,we have demonstrated that the EF1α promoter in conjunction with 2Apeptide sequences could successfully mediate the co-expression of EGXA,BGL4 and XYNB genes in transgenic mice.The study presents a feasible method for generating genetically modified animals with enhanced feed conversion ability.

关 键 词：Β-葡萄糖苷酶 纤维素酶 共表达 转基因小鼠 木聚糖酶 2A肽 

分 类 号：Q78[生物学—分子生物学]