摘要：
In order to study the feasibility of injecting Chinese shrimp (Fenneropenaeus chinensis) total DNA into common carp (Cyprinus carpio) and its effects on muscle nutritional composition of experimental carp,total DNA of Chinese shrimp was fragmented and injected into oosperm of common carp to breed a batch of microinjection offspring.The amplified fragment length polymorphism (AFLP) detection showed that all F2 generation of microinjection contained exogenous gene fragments from the total DNA of Chinese shrimp genome.Furthermore,PCR experiments were conducted using E-AAG and M-CTC primers to detect the Chinese shrimp DNA fragments,which presence in F2 generation of microinjection carp,but absent in control group of carps.Results showed that Chinese shrimp gene fragments had been incorporated into the genome of microinjection carp offspring and can be stably transmitted to the progeny generation.Based on the confirmation of genetic stability of exogenous gene,nutrient content and metal content were tested in the muscle of F2 generation.The resuks showed that crude protein,fat content and ash content in muscle of microinjection carp showed significant difference in that of control carp.Crude protein content was 18.37％ in microinjection carp and 16.49％ in control carp,respectively.Fat content was 2.49％ in microinjection carp,and it was lower than 3.51％ in control carp.Ash content was 1.17％ in microinjection carp,and it was higher than 1.05％ in control carp.Total amino acid content was 79.92％ and 74.16％ in microinjection carp and control carp,respectively.Of which,glutamic acid was the highest amino acid in all the 17 tested amino acids,accounting for 16.60％ and 16.22％ of the total amino acid,with a percentage of 13.27％ and 12.03％ in microinjection carp and control carp,respectively.The top 5 amino acids in microinjection carp were glutamic acid,aspartic acid,lysine,leucine and alanine,which were consistent with that in control carp.These results suggested that there was no differences in amino acid composition between microinjection carp and common carp.The delicious amino acid was 31.16％ in microinjection carp,and it was 28.04％ in control carp,accounting for 38.99％ and 37.81％ of the total amino acid,separately.In addition,tryptophan,isoleucine,leucine,threonine,valine,methionine,phenylalanine and lysine were another 7 essential amino acids in 18 amino acids.The essential amino acid content was 31.5％ and 29.54％ in microinjection carp and control carp,accounting for 39.41％ and 39.83％ of total amino acids,respectively.The limited amino acid was valine in both microinjection carp and control carp.Above all,microinjection carp had certain advantages in nutrients when compared to common carp.Amino acid score (AAS),chemical score (CS) and essential amino acid index (EAAI) were computed by converting the amino acid content into amino acid weight per gram of nitrogen following by the standard evaluation of protein made by FAO/WHO and the amino acid pattern of egg protein.The content of heavy metals including Cu,Zn,Cr,Hg,Cd and Pb in both microinjection carp and common carp met the requirements of relevant food safety standards.This study provides a basis for the commercialization of microinjection carp.%为了研究中国明对虾(Fenneropenaeus chinensis)总DNA直接导入鲤鱼(Cyprinus carpio)后,其对鲤鱼肌肉营养成分的影响,本研究依靠显微介导远缘杂交技术,将远缘基因(中国明对虾基因组DNA)导入鲤鱼受精卵内并获得了子代,对其子代进行扩增片段长度多态性(amplification fragment length polymorphism,AFLP)检测,均扩增出了供体基因片段.在确定了外源DNA稳定遗传的基础上,测定了显微介导中国明对虾基因鲤F2代肌肉中的营养成分、金属元素含量,发现显微介导中国明对虾基因鲤与普通鲤肌肉中粗蛋白质含量、氨基酸总量、4种鲜味氨基酸总含量、必需氨基酸总量,分别为18.37％和16.49％、79.92％和74.16％、31.16％％和28.04％、31.5％和29.54％,均高于普通鲤,两种鲤鱼的第一限制氨基酸都是缬氨酸.与普通鲤相比较,显微介导中国明对虾基因鲤在营养价值上具有一定优势.在所测的显微介导中国明对虾基因鲤和普通鲤的重金属Cu,Zn,Cr、Hg,Cd和Pb含量均符合相关食品安全标准要求.本研究结果为显微介导中国明对虾基因鲤的商品化提供了理论依据.
