Figure 5.

Analyses of quantitative RT-PCR and immunodetection. A, Equal amount of total RNA samples isolated from three different developmental stages of cv Williams 82 soybean leaves were used to quantitatively amplify GmMMP2 fragment. A 26S ribosome fragment was amplified as an internal control. One forward gene-specific primer was used in the amplifications together with a reverse polyT-adaptor primer for the GmMMP2 and 26S ribosome genes. B, Ten micrograms of total soluble protein extract was separated with a 4% to 20% (w/v) gradient SDS-PAGE gel, and immunodetection was performed using rabbit antisera against GmMMP2. Two different-size GmMMP2 forms are presented. Lanes 1 through 3 represent young, mature, and senescent leaves, respectively.