摘要
本研究克隆了驴脂肪酸去饱和酶2(FADS2)基因并进行了生物信息学分析,检测了其在驴不同组织中的表达。通过设计FADS2基因引物,PCR克隆得到其CDS序列,然后对FADS2基因编码产物进行生物信息学预测,同时使用实时荧光定量PCR(RT-qPCR)检测驴心脏、肝脏、脾脏、肺脏、肾脏、肌肉、脂肪、乳腺内FADS2基因相对表达量。结果显示:驴FADS2基因CDS长1335 bp,可编码444个氨基酸,FADS2蛋白分子质量为52.43 ku,等电点为8.29,不稳定指数为37.22,疏水性总平均值为-0.231,属于稳定碱性亲水蛋白;FADS2蛋白二级结构主要由α-螺旋(51.80%)和无规则卷曲(37.61%)构成。FADS2基因在所检测驴组织中均有表达,其中在脂肪和乳腺中相对表达量最高,其次为脾脏、肺脏、肾脏、心脏和肝脏,肌肉中相对表达量最低。结果提示,FADS2可能在驴脂肪和乳腺等组织不饱和脂肪酸的合成过程中发挥重要作用。本研究可为进一步探究驴脂肪和驴乳中高不饱和脂肪酸的合成机理提供基础。
In this study,the donkey fatty acid desaturase 2(FADS2)gene was cloned and its expression in different tissues of the donkey was detected by bioinformatics analysis.The FADS2 gene primers were designed and the CDS sequence of FADS2 gene was obtained by PCR cloning,and then the coding products of FADS2 gene were predicted by bioinformatics.The relative expression levels of FADS2 gene in heart,liver,spleen,lung,kidney,muscle,fat and mammary gland was detected by real-time fluorescent quantitative PCR(RT-qPCR).The results showed that the donkey FADS2 gene’s CDS was 1335 bp which may encode a peptide with 444 amino acid residues;the molecular mass,isoelectric point,instability index,and hydrophobicity average value of FADS2 protein were 52.43 ku,8.29,37.22,and-0.231,respectively,and it was a alkaline hydrophilic protein with an stable secondary structure.The secondary structure of FADS2 protein was mainly composed of α-helix(51.80%)and random coil(37.61%).The FADS2 gene were expressed in all tested tissues of donkeys,and the relative expression level was the highest in fat and mammary gland,followed by spleen,lung,kidney,heart,liver,and the relative expression level in muscle was the lowest.The results suggest that FADS2 plays an important role in the synthesis of unsaturated fatty acids in fat and mammary gland of donkeys.This study provided a basis for further exploring the synthesis mechanism of high unsaturated fatty acids in fat and milk of donkeys.
作者
黄飞
杜心怡
刘桂芹
王长法
周苗苗
HUANG Fei;DU Xinyi;LIU Guiqin;WANG Changfa;ZHOU Miaomiao(Liaocheng Research Institute of Donkey High-Efficiency Breeding and Ecological Feeding,School of Agricultural Science and Engineering,Liaocheng University,Liaocheng 252000,China)
出处
《动物营养学报》
CAS
CSCD
北大核心
2024年第2期1204-1212,共9页
CHINESE JOURNAL OF ANIMAL NUTRITION
基金
国家重点研发计划子课题(2022YFD1600103)
山东省自然科学基金(ZR2023MC149)
山东省重点研发计划(2021TZXD012)
聊城大学畜牧学学科开放课题(319462207-9)。
