Cloning and Expression Analysis of a Cotton Microtubule Associated Protein Gene of GhSP1L5

Abstract

Abstract: A cotton Spiral1-Like5（GhSP1L5）full-length open reading frame（ORF）cDNA was cloned from fast elongating fiber tissue by RT-PCR method, The GhSP1L5 gene contains 315 bp necleotides and codes a protein of 105 amino acids. Through sequence function analysis and homology sequence alignment, GhSP1L5 protein includes several microtubule associated protein binding sites and SCOP domains. Phylogenetic tree analysis showed that GhSP1L5 has a similar relationship with SP1L5 protein family. Prokaryotic expression vector pET28a-GhSP1L5 was constructed and transformated into E.coli BL21（DE3）. Recombinant GhSP1L5 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of ~12 kD. Quantitative RT-PCR analysis showed that the GhSP1L5 gene expresses highly at 15 dpa and 20 dpa fiber tissues compared with the expressions in root, stem, leaf and fiber tissuse of other development stages. GhSP1L5 cloning and expression analysis establish a basis for its further functions in cotton fiber development.

Key words: Cotton fiber, Spiral1-Like5 gene, Microtubule associated protein, Expression

Li Rong, Kou Wei, Tan Lan, He Yi, Liang Zhuo, Li Hongbin. Cloning and Expression Analysis of a Cotton Microtubule Associated Protein Gene of GhSP1L5[J]. Biotechnology Bulletin, 2014, 0(10): 82-87.

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