奶山羊LF基因CDS区克隆、生物信息学分析及其乳中含量测定

doi:10.16431/j.cnki.1671-7236.2018.10.017

摘要/Abstract

摘要：

为了解奶山羊乳铁蛋白（lactoferrin，LF）基因的生物学特性，分析萨能奶山羊不同泌乳时期乳中LF含量的变化，试验扩增了萨能奶山羊乳腺上皮细胞LF基因CDS区序列，运用生物信息学软件对奶山羊LF基因的核苷酸序列特性、编码蛋白的理化性质、保守结构域及高级结构、与其他物种的同源性等进行了分析，并对奶山羊乳样品进行了ELISA测定。结果表明，萨能奶山羊LF基因长为2 127 bp，编码708个氨基酸，编码蛋白质的分子式为C₃₄₀₅H₅₃₆₅N₉₄₉O₁₀₂₄S₄₃，是一个不稳定的亲水性蛋白，二级结构包含α-螺旋、延伸链和无规则卷曲，具有与转铁蛋白相似的折叠方式；羊乳铁蛋白氨基酸残基组成单链，形成77.359 ku具有高级结构的糖蛋白；通过比对发现，牛科LF氨基酸序列的同源性较高（>92%），与其他物种的差异较大，其中萨能奶山羊与山羊的同源性最高（99%），且处于进化树的同一个分支。ELISA检测结果发现，奶山羊初乳中LF含量最高，常乳中的含量较低且较为稳定。研究结果初步表明了萨能奶山羊LF序列和结构域特点，揭示了萨能奶山羊乳中LF含量的变化规律，为LF功能研究、产品研发等提供参考。

关键词: 萨能奶山羊; 乳铁蛋白(LF)基因; 克隆; 生物信息学

Abstract:

In order to explore the biological characteristics of lactoferrin (LF) gene in dairy goat,and analyze the variations of LF of milk in Saanen dairy goats,CDS region of LF gene in Saanen dairy goat was cloned,the sequence,physicochemical property,conservative structure domain and homology comparison of LF gene were analyzed by bioinformatics softwares,milk samples from individual were obtained and determined using ELISA kit.The results showed that the length of LF gene in Saanen dairy goat was 2 127 bp,which coded 708 amino acids.The formula of LF protein was C₃₄₀₅H₅₃₆₅N₉₄₉O₁₀₂₄S₄₃,which was an unstable and hydrophilic protein.The secondary structure of LF mainly contained α-helix,extended chain and random coil,similar with that in transferrin.The molecular weight of LF was 77.359 ku.The protein sequence analysis result showed that the high similarity were found in Bovidae (> 92%),and the rest were quite different,LF gene of Saanen dairy goat exhibited the highest homology with goat,they shared 99% amino acid identity and were in the same branch of an evolutionary tree.ELISA results showed that the maximal concentrations were observed in the colostral samples,the common milk had the lower and stable concentrations.The results obtained the sequence and molecular structure of LF gene,and revealed the change of LF content in milk of Saanen dairy goats,which provided a theoretical basis for LF functional researches and product development.

Key words: Saanen dairy goat; LF gene; cloning; bioinformatics

中图分类号:

王建珏, 史怀平, 马功珍, 黄江涛, 吕明, 张志飞. 奶山羊LF基因CDS区克隆、生物信息学分析及其乳中含量测定[J]. 《中国畜牧兽医》, 2018, 45(10): 2805-2813.

WANG Jianjue, SHI Huaiping, MA Gongzhen, HUANG Jiangtao, LV Ming, ZHANG Zhifei. Cloning and Bioinformatics Analysis of LF Gene CDS Region and Its Content Determination of Milk in Dairy Goats[J]. , 2018, 45(10): 2805-2813.

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