| Autor: |
Ailan Zou, Wenju Zhang, Qinyan Pan, Simei Zhu, Jingjing Yin, Runan Tian, Hongwei Gu, Xiaoming Wang, Jinliang Qi, Yonghua Yang |
| Zdroj: |
Plant Cell, Tissue & Organ Culture; Jul2011, Vol. 106 Issue 1, p71-79, 9p |
| Abstrakt: |
Ethylene is a crucial signal in regulating the biosynthesis of shikonin in Lithospermum erythrorhizon. Arabidopsis ethylene-insensitive 3 (EIN3) is the key transcriptional factor of the ethylene signal transduction pathway; thus, EIN3 homologs might play an important role in shikonin formation. Here, LeEIL- 1, a gene with high similarity to EIN3, was cloned from L. erythrorhizon using a combination method of touch-down PCR and rapid amplification of cDNA ends. The full-length cDNA of LeEIL- 1 is 2,359 bp, encoding a polypeptide of 635 amino acids. By inserting the entire coding region of LeEIL- 1 into the pET32a (+) expression vector, the prokaryotic expression of LeEIL- 1 was successfully induced by isopropyl β- d-1-thiogalactopyranoside in BL21(DE3)pLysS cells. Moreover, the recombinant LeEIL-1 protein was verified by western blotting with the Arabidopsis anti-EIN3 antibody . Real-time PCR results show that the mRNA level of LeEIL- 1 was first dramatically induced within 3 h when the L. erythrorhizon cells were transferred from B5 to M9 medium for shikonin formation; subsequently, the transcripts of LeEIL- 1 decreased to a relatively stable level. Tissue-specific expression analysis indicates that less LeEIL- 1 mRNA accumulated in the stem and leaf than in the root, where shikonin was biosynthesized. These results imply that LeEIL- 1 could be a possible reverse genetic target for revealing the relationship between ethylene and shikonin formation. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
|
Full text from SpringerLink