Cloning of ACC Synthase Gene from Paeonia suffruticosa and Construction of Antisense Plant Expression Vector

doi:10.7668/hbnxb.2010.06.007

Abstract

Abstract: According to the DNA full—length sequence of ACC synthase gene fragment of Paeonia suffruticosa (FJ769773)in GenBank database, a pair of specific primers, P1 and P2, were designed and synthesized.Based on the optimized PCR amplification system and high fidelity DNA polymerase KOD—Plus, ACS gene fragment P_sACS-4 was successfully amplified from the total DNA which was extracted from leaves of peony cuhivar Luo Yang Hong.Sequencing resuh indicated that the length of P_sACS-4 fragment was 970 bp, and the similarity of the sequence of P_sACS-4 and the target sequence was 100%.Two sequences alignment analysis revealed that sequence of P_sACS-4 didn 7t contain any intron sequence.The recombinant plasmid and the plant expression vector pBll21 were digested with the SacI and Sma I digest enzyme.After the fragment of ACC synthase gene and pBll21 were recovered and ligated, P_sACS一4 gene fragment was inserted into 35S promoter downstream of plant expression vector pBll21 reverse in orientation, and an antisense plant expression vector pBll21一anti-PsACS-4 was constructed successfully.

Key words: Paeonia suffruticosa, ACS, Antisense plant expression vector

CLC Number:

S685.11

FAN Bing-you, GAO Shui-ping, LIU Gai-xiu, SHI Guo-an, LI Jia-jue, KONG Xiang-sheng. Cloning of ACC Synthase Gene from Paeonia suffruticosa and Construction of Antisense Plant Expression Vector[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(6): 34-37. doi: 10.7668/hbnxb.2010.06.007.

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