Part:BBa_K2029000:Experience

Using nprb in E. coli with a high-copy backbone such as pSB1C3, is difficult since the constitutive nature of the P43 promotor severely limits cell growth. Colonies took several days to develop in size and the number of colonies was limited. Using a low copy backbone such as pSB4C5, E. coli growth was improved as can be seen by both the speed with which colonies developed (data not shown) as well as the colony number (compare table 1).

Sample	Plate 1	Plate 2
nprb + C3	0	0
nprb + C5	700	300

Table 1. Growth of E.coli bacteria transformed with equal amounts of pP43-nprb in the high-copy backbone pSB1C3 (“C3”) and the low-copy backbone pSB4C5 (“C5”) after 16h of incubation at 37°C.

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