Team:Hong Kong HKUST/riboregulator/characterization

Introduction

In our characterization, comX protein and PcelA (100 base pairs combox promoter) was assembled with GFP reporter (BBa_E0240) using RFC10 format (BBa_K1379005). The construct BBa_K1379005 was then transformed into DH10B cells. To provide a positive control, BBa_I20260 containing strong constitutive promoter, strong RBS, and GFP generator was transformed into DH10B cells. Our negative control, BBa_K1379002, was the same as our experimental construct, but without the comX gene.

To measure the RPU (Relative Promoter Unit) of PcelA promoter, we adopted the method described by Kelly et al. in “Measuring the activity of BioBrick promoters using an in vivo reference standard” (Kelly et al., 2009). With the use of EnVision multilabel reader from Perkin Elmer Company, it enabled us to obtain the fluorescence and absorbance of cells over time. The experimental construct used was BBa_K1379005, which contain comX protein generator, PcelA, and GFP generator. The positive control used in this characterization is BBa_I20260 which is a GFP generator, while the negative control used in this characterization is BBa_K1379002 which is PcelA with GFP and without comX.

Results

Figure 1. PcelA and Phelicase promoter induced by com-X protein drives GFP expression. PcelA and Phelicase promoter induced by com-X protein gave GFP signals. While the same construct without com-X protein did not give any GFP signals. Another negative control which is only protein comX without PcelA/Phelicase also did not give any GFP signals. Reference promoter J23100 + GFP is used as positive control. Scale bar = 5mm.

The meaning of life is a philosophical question concerning the significance of life or existence in general. It can also be expressed in different forms, such as "Why are we here?", "What is life all about?", and "What is the purpose of existence?" It has been the subject of much philosophical, scientific, and theological speculation throughout history. There have been a large number of proposed answers to these questions from many different cultural and ideological backgrounds. The meaning of life is in the philosophical and religious conceptions of existence, social ties, consciousness, and happiness, and borders on many other issues, such as symbolic meaning, ontology, value, purpose, ethics, good and evil, free will, the existence of one or multiple gods, conceptions of God, the soul, and the afterlife.

Fig 1 . Here is the potato.

Here is the description of the potato: it is a potato!

Scientific contributions focus primarily on describing related empirical facts about the universe, exploring the context and parameters concerning the 'how' of life. Science also studies and can provide recommendations for the pursuit of well-being and a related conception of morality.

Fig 1 . Here is the potato.

Here is the description of the potato: it is a potato!

Scientific contributions focus primarily on describing related empirical facts about the universe, exploring the context and parameters concerning the 'how' of life. Science also studies and can provide recommendations for the pursuit of well-being and a related conception of morality.

The meaning of life is a philosophical question concerning the significance of life or existence in general. It can also be expressed in different forms, such as "Why are we here?", "What is life all about?", and "What is the purpose of existence?" It has been the subject of much philosophical, scientific, and theological speculation throughout history. There have been a large number of proposed answers to these questions from many different cultural and ideological backgrounds. The meaning of life is in the philosophical and religious conceptions of existence, social ties, consciousness, and happiness, and borders on many other issues, such as symbolic meaning, ontology, value, purpose, ethics, good and evil, free will, the existence of one or multiple gods, conceptions of God, the soul, and the afterlife.