Product Data Sheet

Recovery

1. Obtain an LB agar plate with the appropriate antibiotic.
2. Using a sterile pipette tip, touch the bacteria growing within the punctured area of the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a sterile pipette tip.)
3. Run this tip lightly over a section of the plate, as shown in the figure, to create streak #1.
4. Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the plate, to create streak #2.
5. Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate, to create streak #3.
6. Grow overnight in a 37 ^o C incubator (unless a different growth temperature is indicated on the plasmid datasheet).
7. In the morning, single colonies should be visible. If the bacterial growth is too dense, re-streak onto a new agar plate to obtain single colonies.

载体信息：

载体描述：

pLacZi is a yeast integration and

reporter vector for use with the MATCHMAKER One-Hybrid System. This plasmid contains the

bacterial lacZ gene downstream of the minimal promoter of the yeast iso-1-cytochrome C gene

(PCYC1). Target elements can be inserted into the MCS upstream of the PCYC1-lacZ reporter. Without

activation from a cis-regulatory element, lacZ expression is very low when the vector is integrated

into the yeast genome. The yeast URA3 gene is used as a selectable marker for integration into the

nonfunctional ura locus of the YM4271 host strain after linearizing the vector at the Nco I or Apa I

site. pLacZi cannot replicate autonomously in yeast. This plasmid contains the ampicillin resistance

gene (Ampr) and the Col E1 origin for selection and propagation in E. coli . Unique restriction sites

are in bold.