真菌Mucor amphibiorum RCS1中一个类S-腺苷-L-高半胱氨酸水解酶基因的蓝光诱导表达  

Blue-light induced expression of S-adenosy-L-homocysteine hydrolase-like gene in Mucor amphibiorum RCS1

作　　者：高雅[1] 王舒[1] 付鸣佳[1] 钟果林[1] 

机构地区：[1]江西师范大学生命科学学院,南昌330022

出　　处：《微生物学报》2013年第9期1001-1008,共8页Acta Microbiologica Sinica

基　　金：国家自然科学基金(31060004;31260010);江西省自然科学基金(2010GZN0115)~~

摘　　要：【目的】确定真菌Mucor amphibiorum RCS1中一个类S-腺苷-L-高半胱氨酸水解酶基因(S-adenosyl-L-homocysteine hydrolase-like,sahhl)受蓝光诱导表达。【方法】以真菌M.amphibiorum RCS1为研究对象,在随机PCR过程中从中获得了一段555 bp长度的DNA序列。以地高辛对这段已知序列进行标记制备探针,通过Northern杂交检测M.amphibiorum RCS1菌丝体培养过程中,由黑暗到蓝光再到黑暗这一光照条件改变的情况下,sahhl基因的转录情况。同时结合应用real-time PCR方法进行分析检测。【结果】经过比对确定这段555bp序列与已经发表的人(Homo sapiens)、家鼠(Mus musculus)和部分真菌的S-腺苷-L-高半胱氨酸水解酶基因sahh有较高的同源性。因此,初步确认这段mRNA序列来自M.amphibiorum RCS1的一个类S-腺苷-L-高半胱氨酸水解酶基因。sahhl基因在黑暗预培养24 h的情况下,蓝光诱导24 h时通过Northern杂交和real-time PCR均可从菌丝体中检测到sahhl基因的大量转录。但sahhl基因在黑暗预培养48 h的情况下,通过real-time PCR没有检测到sahhl基因的大量表达。【结论】上述结果说明,蓝光可以诱导M.amphibiorum RCS1中生长旺盛的菌丝体中sahhl基因的表达。[ Objective] To determine blue-light induced expression of S-adenosyl-L-homocysteine hydrolase-like （sahhl） gene in fungus Mucor amphibiorum RCS1. [ Methods] In the random process of PCR, a sequence of 555 bp was obtained from M. amphibiorum RCS1. The 555 bp sequence was labeled with digoxin to prepare the probe for northern hybridization. By northern hybridization, the transcription of sahhl gene was analyzed in M. amphibiorum RCS1 mycelia culture process from darkness to blue light to darkness. Simultaneously real-time PCR method was used to the sahhl gene expression analysis. [ Results] Compared with the sequence of sahh gene from Homo sapiens, Mus musculus and some fungi species, a high homology of the 555 bp sequence was confirmed. Therefore, the preliminary confirmation has supported that the 555 bp sequence should be sahhl gene from M. amphibiorum RCS1. Under the dark pre-culture in 24 h, a large amounts of transcript of sahhl gene in the mycelia can be detected by northern hybridization and real-time PCR in the condition of 24 h blue light. But a large amounts of transcript of sahhl gene were not found in other detection for the dark pre-culture of 48 h, even though M. amphibiorum RCS1 mycelia were induced by blue light. [ Conclusion] Blue light can induce the expression of sahhl gene in the vigorous growth of M. amphibiorum RCS1 mvcelia.

关 键 词：S-腺苷-L-高半胱氨酸水解酶 蓝光 MUCOR amphibiorum RCS1 NORTHERN杂交 real-time PCR 

分 类 号：Q78[生物学—分子生物学]