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Figure 4.15 A, B
Restriction analysis of tlie plasniids used for tra~lsforniation. A. The plasmid
pRTY9:GUS-lnt (6.9 kb) was tiscd In biolistic method ol'transfomiatio~~. Lane 1 sliows
hDNA digested with UsrE I1 etlzynie as marker. Lanes 2 and 3 liave duplicate plasln~d
un-restricted sample and lane 4 shows plas~iiid a!kr restr~ction with EcuRl showing Sour
fragments. B. Two un-restr~cted plasmid constructs that were used in Agroboc.ierrii~,i-
mzdtated transl'on~iation. Lane I shows hDNA d~gestcd w1i11 tiilid III ellzymc as 111arkcr.
Lalies 2 and 3 llad pHS737:SBTI (14.3 kb) ~plasni~d wl~ile 1,111~s 4 and 5 llad pHS723:Bt
(15 5 kb) plasm~d preparatluns.
Figure 4.16 A - C
GUS histochenlical assay of the leaflets Srum pl~tdtivcly transfornlcd plants 01'
chickpea. A. 4 closer view ol'tlle leaflet sllowing GUS activity in [lie vclns, B, C. GUS
activity as seen 111 tlie pcliole and veins of lallets.
Figure 4.17 A, B
PCK amplificat~on of 700 bp fragnlcnt of 1ipi11 gene fro111 \he gclio~~i~c DNA5 oSTu
generation pla~its rratisSoni~ed w~th Brcr)>lAb and SBTl genes via Agrubricieriut~i-
mediated transfonnalion. A. ~iplll-PCR of pli~nts lrallsforilled w~lli pMS723:Bt via
Agroborieriu~~r-1n2dlatcd trallsfomiation. Lanes 1 to 10 lid transfomled samples and
show the aniplification of ~tpfll gene. Lane I I negative co~ltrol and 12 lo 17 wcrc positive
controls froni plasmid pHS723:Bt used for tralislbrniatio~~. Lane 18 IS - DNA and h
DNA-BsiE I1 m:~rker was dddcd in the lane 19. Ll. ~rprll-PCR or plants lransfomicd \rill1
pHS737:SBTI vector vln Agvobucieriu1?i-11ied1;itcd translorn~at~on. Lanes 1 to 9 had
transformed samples CS5 to CS9. Lonc 10 L\,;IS negative coiitrol and l I
to I5 were
posltlve controls oSpHS737:SBTI vector tlial w;is iised In the transforni;ttion. ?, DNA-
BsiE I1 marker was added in the lalie 16.