金铁锁β-actin基因克隆及其作为内参基因的研究  

Cloning of β-actin Gene in Psammosilene tunicoides and Its Role as a Reference Gene

作　　者：李媛[1] 张爱丽[1] 李国栋[1] 钱子刚[1] 

机构地区：[1]云南中医学院中药材优良种苗繁育工程中心,云南昆明650500

出　　处：《中药材》2016年第9期1971-1974,共4页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金(81260609);云南省教育厅重点项目(2015119);云南省教育厅科学研究基金(2015J101)

摘　　要：目的:建立金铁锁β-actin基因实时荧光定量PCR方法,为金铁锁实时荧光定量PCR的检测提供了内参基因。方法:根据Gen Bank上β-actin基因保守区域设计特异性引物,利用PCR扩增的方法扩增β-actin目的片段。并以β-actin作为内参基因,利用荧光定量PCR技术对糖基转移酶基因(UGT)在金铁锁根、茎、叶组织中的表达情况进行分析。结果:扩增到金铁锁的β-actin基因序列,长度为153 bp,与王不留行、鹅肠菜、马齿苋的β-actin有较高的同源性。而糖基转移酶基因在以β-actin作为内参基因时能够在金铁锁的不同组织稳定表达。结论:金铁锁β-actin基因是一个可靠的内参基因,适合在金铁锁三萜皂苷生物合成相关功能基因的表达研究中作为内参基因。Objective： To establish a real-time quantitative PCR method to detect Psammosilene tunicoides β-actin,and to provide a reference gene for the detection of Psammosilene tunicoides genes by q PCR. Methods： Specific primers were designed based on the conserved region of the β-actin gene（ Gen Bank） and were used to amplify β-actin by PCR. β-actin was also used as a reference gene in the q PCR analysis of glycosyltransferase gene（ UGT） expression in the roots,stems,and leaves of Psammosilene tunicoides. Results： The length of the β-actin gene amplicon from Psammosilene tunicoides was 153 bp and shared relatively high homology with β-actin found in Vaccaria segetalis,Myosoton aquaticum and Portulaca oleracea. Furthermore,UGT was revealed to be stably expressed in different Psammosilene tunicoides tissues when β-actin was employed as the reference gene. Conclusion： β-actin is a reliable and suitable reference gene for studies on the expression of triterpenoid saponin biosynthesis-related genes in Psammosilene tunicoides.

关 键 词：金铁锁 Β-ACTIN 实时荧光定量PCR TapMan探针 

分 类 号：R282.71[医药卫生—中药学]