Sucrose synthetase gene Ac-Sy1 in pineapple fruits, encoded protein and gene cloning method thereof

Abstract

The invention relates to a gene sequence of a sucrose synthetase gene Ac-Sy1 in pineapple fruits and a protein sequence encoded by the same, belonging to the technical of biology. The invention further relates to a cloning method of the gene, which comprises the following steps of: extracting the total RNA in the pineapple fruits by using a modified SDS method; detecting the total RNA in the pineapple fruits by using agarose gel electrophoresis; carrying out reverse transcription to synthesize a first chain cDNA by using the total RNA as a template and using Oligo dT as a primer for; using the cDNA as a template and carrying out PCR amplification by using the Ex-Taq of Takara; connecting a PCR product with a pGEM-T easy Vector, converting, screening the positive for cloning, further validating by PCR and sequencing to obtain the Ac-Sy1 gene. The gene can provide the theoretical evidence for scientifically adjusting and controlling the quality of the pineapple fruits.

Description

technical field

[0001] The present invention relates to a gene and its cloning method, and at the same time relates to the coded protein of the gene, in particular to a pineapple sucrose synthase gene Ac-Sy1, the coded protein and a gene cloning method, which belong to the field of biotechnology, and in particular to the gene research for fruit quality improvement field. Background technique

[0002] Sucrose synthase (SS) is a soluble enzyme that exists in the cytoplasm. It is a tetramer composed of subunits with a molecular weight of 83-100kD. The optimum pH value for the sucrose synthesis direction is 8.0-9.5, and the optimum pH value for the sucrose cracking direction The value is 5.5-6.5, which catalyzes the following reversible reaction: Suc+UDP←→Fru+UDPG, whether it plays a role in synthesis or decomposition in sucrose metabolism is related to whether the enzyme protein is phosphorylated (Amor et al., 1995; Tanase and Yamaki, 2000 ; Tanase et al., 2002). Since sucros...