A primary study of high performance transgenic rice through maize Ubi-1 promoter fusing selective maker gene . | [Pakistan Journal of Botany • 2012]

Author(s):

1. Jinjuan Shen: Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education; Laboratory of Mechano-developmental Biology, Bioengineering College, Chongqing University, Chongqing 400044, China

2. Pingzhong Cai: Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education; Laboratory of Mechano-developmental Biology, Bioengineering College, Chongqing University, Chongqing 400044, China; Institute of Biotechnology and Nuclear Technology, Sichuan Academy of Agricultural Science, Chengdu 610066, China

3. Feng Qing: Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education; Laboratory of Mechano-developmental Biology, Bioengineering College, Chongqing University, Chongqing 400044, China

4. Zhiyong Zhang: Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education; Laboratory of Mechano-developmental Biology, Bioengineering College, Chongqing University, Chongqing 400044, China

5. Guixue Wang: Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education; Laboratory of Mechano-developmental Biology, Bioengineering College, Chongqing University, Chongqing 400044, China

Abstract:

Based on the expression vector pBI121, we successfully constructed a plant overexpression vector of Hspa4 gene fusing with selective maker gene (hygromycin-resistance gene) driven by the Ubi-1 promoter (pBI121-Ubi-Hpt-Hspa4, p121UHH). The plant expression vectors p121UHH and pCAMBIA1301-Ubi-Hspa4 (p1301UH) were transformed into the rice callus, mediated by Agrobacterium tumefaciens. We screened 17 p121UHH-positive transgenic plants and 15 p1301UH-positive transgenic plants by the hygromycin-resistance gene. The pick-up rate of the resistance callus was 51.7% and 42.5%, respectively, and the rate of regeneration for the resistance callus was 51.2% and 49.1%, respectively. The result of polymerase chain reaction (PCR) identification indicated that the pick-up rate of positive transgenic plants was 51.7% and 42.5% and the total transformation efficiency was 16.5% and 6.2%, and the former was 2.66 times of the later. The results of the experiment indicate that the possibility of the appearance of false positive results in the fusing of a plant overexpression vector with a selective maker gene is much less.

Page(s): 501-506

DOI: DOI not available

Published: Journal: Pakistan Journal of Botany, Volume: 44, Issue: 2, Year: 2012

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