[PDF][PDF] Cloning and analysis of defensin 1 cDNA from Scots pine
VA Kovalyova, IT Gout, RG Kiyamova… - Biopolymers & …, 2007 - academia.edu
Biopolymers & Cell, 2007•academia.edu
A pro tein with antifungal ac tiv ity and a mo lec u lar weight of 5.6 kDa was pu ri fied from 7-
day-old Scots pine seed lings. The LC-MS/MS anal y sis re vealed that pu ri fied pro tein is
highly ho mol o gous to plant defensins. The ob tained se quences and da ta base searches
al lowed us to de sign a set of prim ers for mo lec u lar clon ing of a full length cod ing se
quence, cor re spond ing to Scots pine defensin 1 (PsDef1). This was achieved by a PCR ap
proach, us ing Pinus sylvestris сDNA li brary as a tem plate. The spe cific prod uct of PCR …
day-old Scots pine seed lings. The LC-MS/MS anal y sis re vealed that pu ri fied pro tein is
highly ho mol o gous to plant defensins. The ob tained se quences and da ta base searches
al lowed us to de sign a set of prim ers for mo lec u lar clon ing of a full length cod ing se
quence, cor re spond ing to Scots pine defensin 1 (PsDef1). This was achieved by a PCR ap
proach, us ing Pinus sylvestris сDNA li brary as a tem plate. The spe cific prod uct of PCR …
A pro tein with antifungal ac tiv ity and a mo lec u lar weight of 5.6 kDa was pu ri fied from 7-day-old Scots pine seed lings. The LC-MS/MS anal y sis re vealed that pu ri fied pro tein is highly ho mol o gous to plant defensins. The ob tained se quences and da ta base searches al lowed us to de sign a set of prim ers for mo lec u lar clon ing of a full length cod ing se quence, cor re spond ing to Scots pine defensin 1 (PsDef1). This was achieved by a PCR ap proach, us ing Pinus sylvestris сDNA li brary as a tem plate. The spe cific prod uct of PCR am pli fi cation was cloned into рЕТ 23d (+) vec tor and se quenced. Bioinformatic anal y sis of gen er ated se quences showed that the cod ing se quence of PsDef1 cDNA has the po ten tial to en code a pro tein of 83 amino ac ids in length. The first 33 amino ac ids cor re spond to the N-ter mi nal sig nal pep tide, which is re moved af ter process ing. The ma ture pro tein pos sesses con served res i dues which are com mon to all plant defensins. Based on antifungal prop er ties and se quence sim i lar i ties PsDef1 was as signed to group 1 of plant defensins.
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