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The Cloning and Sequence Analysis of Plasma Membrane Intrinsic Protein Gene (BiPIP) and Actin in Bromus Ircutensis Kom

Author: ZhangJiaJia
Tutor: HanBing
School: Inner Mongolia Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: Bromus ircutensis Kom. Aquaporin Actin RT-PCR RACE
CLC: Q943.2
Type: Master's thesis
Year: 2012
Downloads: 15
Quote: 0
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Abstract


Plant aquaporin formed water selective passageway that mediate the water-transme-mbrane transportation of inside the cell or intercellular and adjust water balance in plants, which may play an important role to improve the water physiology integration of clonal plants and the salt, drought resistance. Study to Bromus ircntensis Kom. for material, Full-length cDNA sequence of BiPIPl-1and BiAct respectively ware acquired by RT-PCR and RACE technology. Their structure of sequence of nucleotide and amino acid were analyzed by bioinformatics software. The mainly results as follows:BiPIP1-1full-length cDNA sequence consists of1077bp, which includes the length of54bp of the5’untranslated region, followed by a156bp of3’untranslated region, and finally867bp of the open reading frame encoding288amino acids. Amino acid sequence of BiPIP1-1not only have the MIP family of signal sequence, but also the characteristics of plasma membrane water channel protein signal sequence. Phylogenetic tree constructed showd that BiPIP1-1are PIP1subfamily members, which are very closer in relationship with HvPIP1-1. Tertiary structure of BiPIP1-1contains six transmembrane structure (Ⅰ～Ⅵ, connected by five loops) and carboxyl terminusa and amino terminus. The two NPA boxes (Asn-Pro-Ala) is located in the center of the spiral, forming a two-way transportation through the water. Cloning of plasma membrane instrinsic protein gene in Bromus ircutensis Kom. is helpful for researching aquaporin expression of the molecular level in roots and the influence mechanism of water in the process of physiology integration.BiAct full-length cDNA sequence consists of1310bp, which includes the length of44bp of the5’untranslated region, followed by a135bp of3’untranslated region, and finally1131bp of the open reading frame encoding376amino acids. Amino acid sequence of BiAct have more than one specificity binding site of Actin gene families that containing ATP binding site and gelsolin binding site and profilin binding site. Phylogenetic tree constructed showd that BiAct are very closer in relationship with ZmAct. Cloning of BiAct provide inner control gene for study expression and regulation of other genes in Bromus ircutensis Kom.

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