水牛泌乳期与非泌乳期乳腺组织差异miRNAs的表达模式鉴定及分析

doi:10.16431/j.cnki.1671-7236.2017.11.016

摘要/Abstract

摘要：

为了探求水牛泌乳和非泌乳期关键调控差异表达miRNAs及其表达模式，试验结合Solexa高通量测序和生物信息学分析结果筛选出18个水牛乳腺组织在泌乳期和非泌乳期差异表达的miRNAs，设计、筛选可行的miRNAs反转录及实时荧光定量PCR引物，进行实时荧光定量PCR验证差异表达miRNAs的表达模式。结果发现，miRNA的差异表达模式与测序结果基本一致。泌乳期miRNA-103、miRNA-125a、miRNA-30a-5p和miRNA-148a的表达量均显著高于非泌乳期（P<0.05）；miRNA-29a在非泌乳期表达量显著高于泌乳期（P<0.05）；miRNA-141、miRNA-125b和miRNA-497在泌乳期表达量均高于非泌乳期，但差异不显著（P>0.05）；低丰度的miRNA-490和miRNA-592在泌乳期表达量均显著高于非泌乳期（P<0.05）。Novel-123b在泌乳期和非泌乳期差异不显著（P>0.05），Novel-57在泌乳期比非泌乳期高29.79倍（P<0.01）。本试验结果为后续水牛乳腺研究提供了16个miRNA的可用特异性反转录引物和实时荧光定量PCR引物，miRNA-103、miRNA-125a、miRNA-30a-5p、miRNA-148a、miRNA-29a和Novel-57 6个水牛差异表达miRNAs值得进一步研究。

关键词: 水牛; 泌乳期; 非泌乳期; 乳腺组织; miRNA; 鉴定; 实时荧光定量PCR

Abstract:

In order to explore the differential expression miRNAs and their expression patterns in the control of buffalo in lactation and non-lactation, the miRNAs of 18 buffalo breast tissues differentially expressed in lactation and non-lactation were screened by Solexa high-throughput sequencing and bioinformatics analysis. Viable miRNAs primers for reverse transcription and Real-time quantitative PCR were used to validate the expression patterns of miRNAs. The Real-time quantitative PCR results were basically consistent with sequencing results. The expression levels of miR-103, miRNA-125a, miRNA-30a-5p and miRNA-148a in lactation were significantly higher than those in non-lactation (P<0.05). The expression level of miRNA-29a in non-lactation was higher than that in lactation (P<0.05). The expression levels of miRNA-141, miRNA-125b and miRNA-497 in lactation were higher than those in non-lactation,but the difference was not significant (P>0.05). The low-expression levels of miRNA-490 and miRNA-592 in lactation were significantly higher than those in non-lactation (P<0.05). There was no significant difference in the expression of Novel-123b between lactation and non-lactation (P>0.05). Novel-57 was extremely significantly in lactation (29.79 times) higher than non-lactation (P<0.01). This results provide available specific reverse transcription primers and Real-time quantitative PCR primers of 16 miRNAs for further study on buffalo,and 6 miRNAs such as miRNA-103, miRNA-125a, miRNA-30a-5p, miRNA-148a,miRNA-29a and Novel-57 deserve further research.

Key words: buffalo; lactation; non-lactation; mammary gland; miRNA; identification; Real-time quantitative PCR

中图分类号:

S814.8

蔡小艳, 王鹏程, 邓凯, 王萍, 冯万有, 张晓溪, 任艳萍, 刘庆友, 石德顺. 水牛泌乳期与非泌乳期乳腺组织差异miRNAs的表达模式鉴定及分析[J]. 《中国畜牧兽医》, 2017, 44(11): 3220-3228.

CAI Xiao-yan, WANG Peng-cheng, DENG Kai, WANG Ping, FENG Wan-you, ZHANG Xiao-xi, REN Yan-ping, LIU Qing-you, SHI De-shun. Identification and Analysis of Differential Expression miRNAs in the Lactation and Non-lactation of Buffalo Mammary Gland[J]. , 2017, 44(11): 3220-3228.

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