用长春优质肉种公牛繁育中心的33头种公牛,通过PCR-SSCP技术,检测细胞色素氧化酶(Cytochrome c ox-idase,COX)中存在的突变点,并对多态片段进行了测序,以确定突变的碱基类型和突变点的位置。根据PCR-SS-CP检测及测序结果得知:在细胞色素氧化酶基因的外显子中,第2外显子的92bp位处存在T→C的突变,该碱基位于密码子第2位,氨基酸由异亮氨酸突变为苏氨酸。同时结合精液品质测定记录,对长春优质肉种公牛繁育中心的33头种公牛的精液品质进行了相关分析,结果显示:细胞色素氧化酶基因的第二外显子CC型的活精子比率与CD型相比存在极显著差异(P<0.01),冻精活力存在显著差异(P<0.05)。
[Objective] The research aimed to discuss the differential expression quantity of miRNA-181b in mature（18-month-old） and immature（one-month-old） cattle＇s anterior pituitary and its regulation function.[Method] cDNA library of miRNA in mature（18-month-old） and immature（one-month-old） cattle＇s anterior pituitary were established.After Solexa high-throughput sequencing of miRNA in the cDNA library,miRNA in anterior pituitary of bulls was identified.miRNA-181b with differential expression were selected from the sequencing results.By real-time quantitative RT-PCR,the expression laws of miRNA-181b in the anterior pituitary of Yanbian Cattle in different growth period was validated.And the target genes of miRNA-181b were forecast by using TargetScanS prediction software.[Result] The expression quantity of miRNA-181b had great difference in cattle＇s anterior pituitary different growth periods.The expression quantity of miRNA-181b in anterior pituitary of one-month-old cattle was 4.05 times as that in 18-month-old cattle.The binding of miRNA-181b with 838-844 bases in 3＇ untranslated region of FSHβ gene was specific and the binding base sites were UGAAUGUA.[Conclusion] This research provided the theoretical basis for the transcription regulation research of FSHβ.
