利用RNA-seq技术和CNVs筛选控制多羔肉羊关键基因及调控网络研究

Research and selecting key genes and molecular regulation mechanism of sheep fecundity traits to improve productivity of mutton sheep is of great significance. It is an effective method to study on key genes and molecular genetics, discover fecundity traits molecular marks and signal regulatory network by RNA-Seq and CNVs tecnology. The project will use producing-multiple-lambs Bamei mutton sheep as test group, and use producing-single-lamb Bamei mutton sheep and hormone treated sheep as control groups, to carry out studies on genome copy number variation and differences in gene expression in reproductive axis. Based on the corresponding relationship between the variation in gene structure and expression differences, select key genes that control sheep prolificacy traits, and research the genetic variation, expression rules of these key genes in different reproductive axis tissues as well as their relationship with lambing number in order to reveal the underlying causes for differences in sheep reproductive from the transcriptome and genome levels. At the same time, we will build gene interaction network for differentially expressed genes using bioinformatic technologies, initially reveal local molecular interactions mechanism of reproductive axis differentially expressed genes. The present study can provide a theoretical basis for protection and exploitation of excellent breeds.

研究和筛选控制肉羊多羔性状的关键基因及其分子调控机理，对提高肉用绵羊生产水平具有重要意义，采用RNA-Seq和基因组拷贝数变异技术开展巴美肉羊多羔关键基因和分子遗传研究，揭示多羔性状作用的关键基因、分子标记和信号调控网络是一项有效的方法。本项目以连续产多羔的巴美肉羊为试验组，以连续产单羔巴美肉羊和激素处理组为对照组，开展基因组拷贝数变异和生殖轴差异表达基因研究，以基因结构变异与表达差异对应关系为依据，筛选出调控巴美肉羊多羔性状的关键基因，研究关键基因在不同繁殖力绵羊的遗传变异、表达规律及其与产羔数的关系等，从转录组和基因组水平揭示绵羊繁殖性状差异的内在原因。同时利用生物信息学技术对差异表达基因构建基因互作网络，初步揭示生殖轴基因表达调控的局部分子互作机制。本研究成果可为优良品种的保护和开发利用提供理论依据。

研究和筛选肉羊多羔性状关键基因及分子调控机理，对提高肉用绵羊生产水平具有重要意义，采用RNA-Seq和基因组拷贝数变异技术开展巴美肉羊多羔关键基因和分子遗传研究，揭示多羔性状作用的关键基因、分子标记和信号调控网络是一项有效的方法。.本研究分析了生殖激素FSH、LH、E2和P4在单、双羔巴美肉羊发情期外周血中的分泌规律和特点，表明了巴美肉羊产羔数增加可能与血清FSH和LH浓度升高有关，探讨了生殖激素与产羔数的相互关系。完成了单、双羔巴美肉羊卵巢差异表达miRNA的筛选，获得了11个差异表达的miRNAs，推测miR-376b-5p及其靶基因IGF-1、DAZL、MTOR和miR-1185-5p及其靶基因AHR可能在调控绵羊繁殖过程中发挥重要的调控作用。.建立了绵羊下丘脑、垂体、卵巢生殖轴相关基因表达谱分析的RNA-Seq分析流程，确定了85个基因为性腺轴上调控排卵以及产羔方面的关键功能基因；完成了单、双羔巴美肉羊重测序CNVs分析，获得了4881个CNV区域，138个区域在单、双羔组中存在差异，涉及38个基因，有30个基因在巴美肉羊下丘脑、垂体、卵巢中表达；整合不同产羔数巴美肉羊性腺轴转录组与重测序CNVs数据，构建基因共表达网络，确定了ACKR1、BCL3、CALB2为影响绵羊多羔性状的核心功能基因。.另外，本研究还对3个核心功能基因表达、结构及功能进行了分析和预测，为进一步研究这3个基因的作用机制奠定了理论基础。