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Molecular Analysis of OsLEA4 and Its Contributions to Improve E. coli Viability

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Abstract

OsLEA4, a late embryogenesis abundant (LEA) protein gene from rice (Oryza sativa L.), contains a 312-bp open reading frame encoding a putative polypeptide of 103 amino acids with a calculated molecular mass of 11.19 kDa and a theoretical pI of 10.04. OsLEA4 polypeptide is rich in Ala (22%), Lys (15%), Glu (9%), His (8%), Thr (8%), and Arg (7%) and lacking in Trp, Cys, Asn, and Phe residues. OsLEA4 protein contains a Pfam:LEA_1 domain architecture at positions 1–73 with three α-helical domains and without β-sheet domain. In silico predictions showed that OsLEA4 protein was strongly hydrophilic with the grand average of hydropathy value of −0.816 and instability index of 27.31. The hydrophilic regions were found in the conserved motif of OsLEA4. OsLEA4 gene was introduced into Escherichia coli, and a fusion protein (∼29.4 kDa) was expressed after isopropylthio-β-d-galactoside inducting by sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis. OsLEA4 protein enhanced the tolerance of E. coli recombinant to high salinity, heat, freezing, and UV radiation, which suggested that OsLEA4 protein may play a protective role under stressed conditions. This is the first successful use of E. coli as a prokaryotic system for LEA production from rice.

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Abbreviations

GRAVY:

Grand average of hydropathy

IPTG:

Isopropylthio-β-d-galactoside

LEA:

Late embryogenesis abundant

LB:

Luria–Bertani

ORF:

Open reading frame

ROS:

Reactive oxygen species

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

UV:

Ultraviolet

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Acknowledgments

This study was supported by the China National Transgenic Major Program (2009ZX08009-109B), the University Innovation Team Construction Project of Chongqing Education Committee (201040), and The Natural Science Foundation Project of Chongqing Education Committee (KJ111108).

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Authors and Affiliations

  1. Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, People’s Republic of China

    Tingzhang Hu, Hua Zeng, Shuai He & Guixue Wang

  2. School of Life Science and Engineering, Chongqing Three Gorges University, Chongqing, 404100, People’s Republic of China

    Tingzhang Hu, Yingmei Wu & Xiaoyun Huang

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  1. Tingzhang Hu
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  2. Hua Zeng
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  3. Shuai He
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  5. Guixue Wang
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  6. Xiaoyun Huang
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Correspondence to Tingzhang Hu.

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Table S1

The GenBank accession numbers of proteins used for analysis (DOC 51 kb)

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Hu, T., Zeng, H., He, S. et al. Molecular Analysis of OsLEA4 and Its Contributions to Improve E. coli Viability. Appl Biochem Biotechnol 166, 222–233 (2012). https://doi.org/10.1007/s12010-011-9418-5

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