Cloning and sequence analysis of three Actin gene fragments from Pseudostellaria heterophylla
10.7501/j.issn.0253-2670.2016.11.021
- Author:
Ling DING
1
Author Information
1. Guiyang College of Traditional Chinese Medicine
- Publication Type:Journal Article
- Keywords:
Actin;
Gene cloning;
Pseudostellaria heterophylla (Miq.) Pax;
RT-PCR;
Sequence analysis
- From:
Chinese Traditional and Herbal Drugs
2016;47(11):1935-1942
- CountryChina
- Language:Chinese
-
Abstract:
Objective: Cloning and sequence analysis of the cDNA fragments encoding Actin gene from Pseudostellaria heterophylla. Methods: Degenerate primers were designed based on the conservated sequences of the cloned Actin gene from other plant species. The core fragments of Actin gene were obtained by reverse transcription polymerase chain reaction (RT-PCR) and the flanking fragments were cloned by using suppression PCR. The expression profiles of Actin gene in different cultivated provenances, organs, and development stages were analyzed by semiquantitative PCR. Results: The three Actin genes obtained were designated PhACT1, PhACT2, and PhACT3, which were extended to 1008, 1008, and 975 bp through suppression PCR, encoding 336, 336, and 325 amino acids, respecifively. The semiquantitative PCR analysis showed that PhACT2 and PhACT3 had a stable expression except PhACT1.Conclusion: Three Actin genes are cloned and the PhACT2 could be used as an internal standard gene for the expression analysis of the functional genes in P. heterophylla.
