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

Abstract

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.