Novel Ti plasmid pElrLCG for saving plant embryos from lethal mutation and its preparation method

Abstract

The present invention discloses a novel Ti plasmid pElrLCG for saving embryos from lethal mutation and a preparation method of the Ti plasmid pElrLCG. Connect 5.211kb long bearer fragment recovered from enzyme cutting of original bearer pJawoh18 with synthesized XhoI-loxP-NheI-NcoI-Nos-BsiwI-KpnI-StuI-BgIII-MFeI-AatII-1xMyc-loxP-SpeI fragment, and then insert Gr-Cre, Gateway Cassettee fragment between two loxP loca, to construct a new plasmid vector pElrLCG. The plasmid vector can be used to obtain homozygote of embryonic lethal mutant or study the development of homozygote of embryonic lethal mutant in late stage, and therefore is of great significance for study on lethal mutations of embryos, hybridization breeding, and special biologic behaviors of lethal mutants of embryos.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a novel Ti plasmid for rescuing lethal mutations of plant embryos: pElrLCG and a preparation method thereof. Background technique [0002] The existing inducible vectors that can be used to rescue lethal mutations in plant embryos are: (1) tetracycline-inducible vectors (2) glucocorticoid-inducible vectors (3) estrogen-inducible vectors (4) alcohol-inducible vectors (5) Ecdysone hormone-inducible carrier (6) copper-inducible carrier, etc. The use of tetracycline-inducible carriers is potentially dangerous to the environment. In addition, there must be a large amount of tetracycline in the nucleus to work, and high concentrations of tetracycline are toxic; the disadvantages of estrogen-inducible carriers are: strong local induction, estrogen in It is not easy to spread in the plant, so the whole plant cannot be induced; the main disadvantage of the ecdysone-inducibl...

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Problems solved by technology

The use of tetracycline-inducible carriers is potentially dangerous to the environment. In addition, there must be a large amount of tetracycline in the nucleus to work, and high concentrations of tetracycline are toxic; the disadvantages of estrogen-inducible carriers are: strong local induction, estrogen in It is not easy to spread in the plant, so the whole plant cannot be induced; the main disadvantage of the ecdysone-inducible carrier is: the leaves have poor absorption of the ecdysone; and the alcohol-inducible carrier is toxic to the plant, and the alcohol is volatile; the copper-inducible carrier The disadvantages of the expression vector are: copper can only work in some cells and is toxic to the plant; the inducing chemical substance of the glucocorticoid-inducible expression vector: dexamethasone, can be efficiently transported to various tissues and organs of the plant and has a negative effect on the plant. The plant is non-toxic, which is the advantage of this carrier

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