【摘要】 旨在鉴定分析水牛乳腺泌乳期和非泌乳期miRNA的表达谱和差异作用机制。采集水牛泌乳期和非泌乳期乳腺组织,利用Solexa测序技术构建2个时期miRNAs表达谱,鉴定前体miRNAs、成熟miRNAs和新miRNAs,分析miRNAs的第一偏好性核苷酸和染色体分布,发掘水牛泌乳期和非泌乳期高表达及差异表达miRNAs。结果显示,本研究构建了水牛泌乳期和非泌乳期乳腺组织2个miRNA表达谱,分别获得12 768 110和12 569 467条18~31nt的高质量序列。测序确认了归属259个miRNAs家族的359个成熟miRNAs和363个pre-miRNAs及5个水牛特有的miRNAs。U是19和25nt miRNAs的5′端最普遍的核苷酸。bbu-let-7b、bbu-let-7a、miR-26a和miR-21等miRNAs在2个时期均呈现高表达;bbu-miR-148a、143、200c、200a和bbu-let-7f等miRNAs在非泌乳期特异性高表达。bbu-miR-125b、29a和bbu-let-7c等miRNAs在泌乳期特异性高表达。bbu-miR-148a、143、200a、141和30a-5p等miRNAs在泌乳期的表达量下降为非泌乳期的1/2以下。而bbu-miR-26a、29a、125b、99a和bbulet-7c等miRNAs在泌乳期表达量大于或等于2倍非泌乳期丰度。本研究构建了水牛泌乳期以及非泌乳期乳腺组织2个miRNAs表达谱,测序确认了259个水牛miRNAs家族的359个成熟miRNAs和363个前体miRNAs,5个水牛特有miRNAs和10个高差异表达miRNAs,为进一步阐明奶水牛泌乳关键miRNAs作用机制奠定了基础。更多还原

【Abstract】 This study aimed to identify and analyze the expression patterns and mechanism of buffalo mammary gland miRNAs in the lactation and non-lactation periods.Buffalo mammary gland tissues in lactation and non-lactation periods were collected,their miRNAs expression profiles were analyzed by Solexa sequencing technology,pre-miRNAs,mature miRNAs and novel miRNAs were identified,respectively,the first preferred nucleotide and chromosome distribution were analyzed.The highly and differentially expressed miRNAs in the lactation and non-lactation periods were detected.The results showed that 2miRNAs expression profiles from buffalo lacta-tion and non-lactation mammary glands were constructed.12 768 110 and 12 569 467high-quality reads between 18 and 31nt were obtained seperately.Three hundred fifty-nine mature miRNAs and 363pre-miRNAs belonged to 259 miRNAs families,and 5buffalo-specific miRNAs were confirmed by sequencing.U was the most common nucleotide at the 5′end of 19 and 25nt miRNAs.The bbu-let-7b,bbu-let-7a,miR-26 aand miR-21 showed high expression in both periods.bbumiR-148 a,143,200 c,200aand bbu-let-7fwere specificially highly expressed in non-lactation period,the bbu-miR-125 b,29aand bbu-let-7cwere specifically highly expressed in lactation period.The bbu-miR-148 a,143,200 a,141and 30a-5p decreased the expression in the lactation period to less than 1/2of the non-lactation period,bbu-miR-26 a,29a,125 b,99aand bbu-let-7cwere highly expressed in the lactation period more than or equal to 2times of the lactation period.The result indicate that 2miRNAs profiles from buffalo lactation and non-lactation mammary gland tissues were constructed.359 mature miRNAs and 363pre-miRNAs belonging to 259 buffalo miRNAs families were identified.5buffalo genome-specific miRNAs and 10 highly differentially expressed miRNAs were obtained,which lay the foundation for further elucidating the mechanism of key miRNAs in lactating buffalo.更多还原