Cloning And Sequences Analysis Of Seabuckthorn Chitinase Gene

Hippophae rhamnoides is an excellent plant for soil and water conservation, wind breaking and sand fixation. But from of old, plant diseases and insect pests are severe. Now traditional methods could not prevent effectively, cultivating a kind of keeping resistance to diseases and insect pests becomes a main way.Chitin is a major constituent of pathogenic fungal cell wall and pest crust. Chitinase can destroy and destruct chitin, then lead pathogeny and pests dead. As a kind of PR protein, chitinase genes have extensive physiologic activities. So it supports us cultiving through transgene method in theoretically.In this study, Hippophae rhamnoides was used as the experimental material. We isolated some pathogeny fungals and inducted to ensure the right pathogeny fungal. We isolated chitinase protein for the first time, and proved the characteristic of resistant fungal. Using PT-PCR and inverse PCR, ClassIII and ClassI chitinase of Hippophae rhamnoides was coloned; sequenced and analyzed.The PI of ClassIII chitinase of Hippophae rhamnoides is 5.90; the molecular weight is 31.758KD; the AAs includes chitinase family 18 active site, 144-152 is the active site, with sequence of LDGIDFDIE; E(Glu) is the active site residue; the raw cleavage site is A-G.. The PI of ClassI chitinase of Hippophae rhamnoides is 8.45; the molecular weight is 32.293KD; the AAs includes one chitin-binding protein from 20-61, with the sequences of chitin-binding protein AEQCGGQAGGRVCRVCPGG ACCSKFVRCGNTADYCGSGCQSQCSS. And we construct their binary structure and 3D-structure respectivly.