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当前位置: 首页 > 文章 > 荣昌猪唾液腺全长cDNA文库的构建及ESTs分析 西北农林科技大学学报(自然科学版) 2017,45 (3) 17-23
Position: Home > Articles > Construction of full-length cDNA library and analysis of expressed sequence tags(ESTs)of salivary glands of Rongchang pig Journal of Northwest A & F University(Natural Science Edition) 2017,45 (3) 17-23

荣昌猪唾液腺全长cDNA文库的构建及ESTs分析

作  者：
甘玲;梅宏远;杨滨宇
单  位：
西南大学荣昌校区动物医学系
关键词：
荣昌猪;唾液腺;全长cDNA文库;LD-PCR;SMART技术;表达序列标签
摘  要：
【目的】构建荣昌猪唾液腺全长cDNA文库,并进行部分表达序列标签(ESTs)的测序及分析,为荣昌猪种质资源的保存、开发和挖掘提供重要的分子信息。【方法】以14日龄荣昌猪仔猪为材料,取其唾液腺,采用SMART法构建唾液腺全长cDNA文库,并进行ESTs测序、注释及功能分析。【结果】初级文库滴度为5.88×106PFU/mL,重组率为99.67%,大部分插入片段长度为400~2 000bp。共获得144条有效ESTs序列,共计108条非重复序列,包括23个重叠群和85个单一序列,77(占71.29%)条序列为已知功能基因,31(占28.71%)条为未知功能序列,其中18条为新基因。GO分类和KEGG途径分析结果显示,荣昌猪唾液腺表达基因功能主要包括细胞成分、分子功能和生物学过程3个类群。在细胞成分类群中,细胞外的成分所占比例最高;在分子功能类群中,酶抑制剂活性所占比例最高;而在生物过程类群中,以转运和定位等过程最突出。表达的基因主要参与补足物和凝固级联、百日咳、加压素调控的水再吸收及味觉传导等代谢途径。【结论】成功构建了荣昌猪唾液腺全长cDNA文库,解析了荣昌猪唾液腺的表达特征。
译  名：
Construction of full-length cDNA library and analysis of expressed sequence tags(ESTs)of salivary glands of Rongchang pig
作  者：
GAN Ling;MEI Hongyuan;YANG Binyu;Department of Veterinary Medicine,Southwest University,Rongchang Campus;
关键词：
Rongchang piglets;;salivary gland;;full-length cDNA library;;LD-PCR;;SMART technology;;expressed sequence tags(ESTs)
摘  要：
【Objective】This study constructed the full-length cDNA library of Rongchang pig salivary glands and conducted the sequencing and analysis on expressed sequence tags(ESTs)to provide molecular information for storing,exploring and digging genetic resources of Rongchang pig.【Method】Salivary glands of Rongchang piglets at the age of 14 days were collected and SMART technology was used to construct the full-length cDNA library.Sequencing,annotation and functional analysis of ESTs were also conducted.【Result】The titer of primary library was 5.88×106 PFU/mL and the ratio of recombinants was99.67%.The average length of most exogenous inserts was among 400-2 000 bp.A total of 144 effective ESTs were obtained,including 108 unigenes with 23 contigs and 85 single sequences.Among these,77(71.29%)sequences were known functional genes,31(28.71%)sequences were unknown,and 18 were novel genes.Gene ontology and KEGG pathways analysis revealed that the functions of genes expressed in Rongchang pig salivary glands mainly included cellular components,molecular functions and biological processes.In the cell component category,the extracellular component had the highest ratio.In the molecular functional category,the inhibitor activity had the highest ratio.Transporting and positioning processeswere the most outstanding in the biological process category.These expressed genes involved in complement and coagulation cascades,pertussis,vasopressin-regulated water reabsorption and taste transduction.【Conclusion】This study successfully constructed the full-length cDNA library and displayed the expression characteristics of Rongchang pig salivary glands.

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