Method for cloning complete sequence of coding region of homologous gene of LFY in plum blossom

Abstract

The invention discloses a method for cloning the complete sequence of the coding region of the homologous gene of LFY in plum blossom. The method comprises the following steps: carrying out genome level cloning; extracting DNA in plum leaves; carrying out primer designing and a PCR reaction; designing a pair of degenerate primers by designing an upstream primer in the upstream region of an initiator and by designing a downstream primer in the downstream region of a terminator codon; recovering and purifying PCR products; cloning; carrying out gene coding region complete-sequence cloning; extracting total RNA in the plum leaves, and carrying out inverse transcription of the total RNA to form cDNA; carrying out primer designing and a PCR reaction: designing a pair of specific primers on the basis of a genome sequencing result, wherein an upstream primer and a downstream primer comprise an initiator and a terminator codon respectively; recovering and purifying PCR products; and cloning. The method has the characteristics of cost saving, simplicity and feasibility, and is suitable for the wide application.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to a method for cloning the complete sequence of the LFY homologous gene coding region in plum blossoms. Background technique [0002] Some studies have shown that the LFY gene is an essential gene that determines the formation of floral meristems (Weigel and N, 1995), and plays a central role in the process of plant flowering. The LFY gene is found in all land plants and has an evolutionary history of nearly 40 million years. In 1990, Coen et al. isolated the homologous gene FLO of LFY from snapdragon grass by transposon tagging method, and found that it plays an important role in the formation of floral meristem. In 1992, Weigel et al. successfully cloned the full length of the LFY gene from Arabidopsis thaliana by studying the lfy1 mutant by using RFLP technology. Through the study of its expression mode and transgene, it was believed that the LFY gene not only controls the ...

AI Technical Summary

Problems solved by technology

There are some defects: 1) The gene fragment length of the target PCR product is not clear; 2) The test cost is high; 3) The workload is heavy and the experimental technology requirements are high

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