What question did this study set out to answer?

Design modified hygromycin resistance gene cassettes with removed internal restriction enzyme sites to facilitate cloning and vector construction for plant transformation.

October 1, 1991PLANT PHYSIOLOGYOpen Access

Hygromycin Resistance Gene Cassettes for Vector Construction and Selection of Transformed Rice Protoplasts

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Authors

ZZZhenwei ZhengRutgers, The State University of New Jersey AHAkio HayashimotoLouisiana State University ZLZhijian LiBroad Institute

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Implication

Laboratory study demonstrates modified hygromycin resistance cassettes enable effective selection in rice protoplasts, suggesting simplified vector construction for plant transformation.

Key Points

Design modified hygromycin resistance gene cassettes with removed internal restriction enzyme sites to facilitate cloning and vector construction for plant transformation.
Mutated the Escherichia coli hygromycin B phosphotransferase (hph) coding sequence to eliminate unique EcoRI, PstI, and SacII restriction sites.
Assembled gene cassettes flanked by multiple cloning sites containing EcoRI, SacII, KpnI, and HindIII.
Assayed hygromycin resistance phenotypes in transformed E. coli and rice protoplast cell cultures.
Engineered hph gene cassettes successfully eliminated internal EcoRI, PstI, and SacII restriction sites without altering coding efficiency.
Mutated hph cassettes conferred hygromycin resistance that was qualitatively indistinguishable from wild-type hph in both E. coli and rice protoplast growth assays.

Cite This Study

Zheng et al. (1991) studied this question.

synapsesocial.com/papers/6aa2ba40d081bdd435a730bd https://doi.org/10.1104/pp.97.2.832

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Implication

Laboratory study demonstrates modified hygromycin resistance cassettes enable effective selection in rice protoplasts, suggesting simplified vector construction for plant transformation.

Hygromycin Resistance Gene Cassettes for Vector Construction and Selection of Transformed Rice Protoplasts

Discussion

Member takes

Implication

Key Points

Cite This Study

Hygromycin Resistance Gene Cassettes for Vector Construction and Selection of Transformed Rice Protoplasts

Discussion

Member takes

Implication

Key Points

Cite This Study

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