Cloning Of The Human Augmenter Of Liver Regeneration Gene CDNA And Transforming Cucumis Melo L.cv Hetao

The 471bp cDNA fragment of human augmenter of liver regeneration (hALR) was amplified from the human fetal liver total RNA by RT-PCR with artificially synthesized two specific primers, then ligated into vector pMD18-T to construct pMD-ALR. The sequence analysis of pMD-ALR showed that it is completely identical with the sequence of hALR cDNA(AF146394) in GenBank. The fruit-specific expression vector pPZP-CAN was constructed by inserting hALR, which was obtained from pMD-ALR by Hindlll and Sac I restrictive digesting, into plant expression vector pPZP-CGN which contains the Cucumis melo L. cv cucumisin gene promoter. Then the pPZP-CAN was introduced into self-pollinating Cucumis melo L. cv Hetao by pollen tube pathway transformation method in testing-farm, and transformed seeds were harvested. PCR, PCR-southern and Dot boltting analysis of seeding showed that hALR have been integrated into the genome of some seeds.These seeds will be further tested for production of hALR protein in...