利用三代全长转录组测序对禽内源性白血病病毒ALVE3不同插入类型鸡进行血液转录组分析  

Blood Transcriptome Analysis for Chickens with Different Types of Insertion Site of Endogenous Avian Leukosis Virus ALVE3 Based on Third Generation Full-length Transcriptome Sequencing

作　　者：王沁园 袁一铭 郑钢 吴俊锋 陈佳华 杨玉琴 杜玉双 连玲[1] WANG Qinyuan;YUAN Yiming;ZHENG Gang;WU Junfeng;CHEN Jiahua;YANG Yuqin;DU Yushuang;LIAN Ling(College of Animal Science and Technology,China Agricultural University,Beijing 100193)

机构地区：[1]中国农业大学动物科学技术学院,北京100193

出　　处：《中国家禽》2024年第10期21-31,共11页China Poultry

基　　金：国家自然科学基金(32272865);国家重点研发项目(2021YFD1300600);国家蛋鸡产业技术体系(CARS-40)。

摘　　要：为进一步探索不同内源性禽白血病病毒3(Avian leukosis virus subgroup E,ALVE3)插入个体间的转录本差异,研究利用PacBio三代测序平台对ALVE3无插入、杂合插入和纯合插入的白来航公鸡血液样本进行全长转录组测序,分别获得27 784、38 460、44 487条转录本,平均长度分别为2 850 bp、3 078 bp和3 166 bp,辅助二代测序数据对其进行校正和定量,并将所得数据与GRCg6a参考基因组比对,将比对到基因组上的转录本进行差异表达分析及lncRNA预测。结果显示,在有ALVE3插入个体中特异存在的转录本有3 657条,将这些转录本利用BLAST与ALVE3全长序列比对,发现转录本Novelgene1474_novel01与ALVE3互补链对应序列高度同源,长度为5 575 bp,说明ALVE3互补链能够产生转录本,对其翻译潜能进行预测后,推测该转录本为lncRNA。此外对不同ALVE3插入情况个体间的差异表达基因进行富集分析,在GO/KEGG中富集到多条免疫相关通路,说明ALVE3的插入可能对机体内免疫相关基因的表达有影响。研究表明,检测到ALVE3反义链可转录产生一段长为5 575 bp的lncRNA,该转录本在介导先天免疫通路中可能发挥作用。To further clarify the transcript of avian leukosis virus subgroup E3(ALVE3).Based on the PacBio sequencing platform,our study used three blood samples of White Leghorn chickens with different types of ALVE3 insertion,including ALVE3-/-,ALVE3+/-and ALVE3+/+.The results showed that a total of 27784,38460 and 44487 transcripts were obtained in ALVE3-/-,ALVE3+/-and ALVE3+/+by sequencing,with an average length of 2850 bp,3078 bp and 3166 bp,respectively.Corrected and quantified by the reads from the next-generation sequencing data,transcripts from the third-generation sequencing were compared to the GRCg6a genome and then performed differential expression analysis and lncRNA prediction.The results showed that there were 3657 transcripts present only in individuals with ALVE3 insertion.These transcripts were BLAST to ALVE3,and Novelgene1474_Novel01 was highly similar with the complementary chains of ALVE3.This study predicted the translation potential of Novelgene1474_Novel01,and we speculated that Novelgene1474_Novel01 was a lncRNA with the length of 5575 bp.In addition,enrichment analysis was conducted on differentially expressed genes among individuals with different types of ALVE3 insertion,and multiple immune-related pathways were significantly enriched in GO/KEGG,indicating that ALVE3 insertion might affect the expression of immune-related genes.Collectively,the results found that the ALVE3 antisense strand could transcribe a lncRNA with the length of 5575bp,which might play a role in mediating the innate immune-related pathway.

关 键 词：鸡 内源性禽白血病病毒 全长转录组测序 

分 类 号：S831.2[农业科学—畜牧学]