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pHSP70/pRBCS2 combined promoter for C. reinhardtii

Improved part from 2016

This part is an improved version of USP_UNIFESP-Brazil 2016 iGEM team. Link: https://parts.igem.org/Part:BBa_K2136013

The combined constitutive endogenous promoter used for foreign gene expression in C. reinhardtii.

This promoter consists of 5 main parts:

1) HSP70A promoter (1-263) is the first upstream region of Heat Shock Protein 70A from C. reinhardtii. This sequence spacing-dependent and acts as an inhibitor of transcriptional silencing in C. reinhardtii. [1]

2) [Improvement] High-efficiency separator sequence between two promoters (264-272) that provides the best spacing between two promoters, resulting in an increase of transformation rate by 2.6 times relative to original separating sequence. [1]

3) RBCS2 promoter (273-454) is a promoter region of nuclear ribulose bisphosphate carboxylase/oxygenase small subunit gene from C. reinhardtii. It is the most widely utilized promoter for robust transgene expression in C. reinhardtii. [1]

4) 5' untranslated region of the nuclear RBCS2 gene (455-489) from C. reinhardtii acts as an enhancer.

5) [Improvement] The first intron of RBCS2 gene (490-634), it acts as an enhancer and increases expression by 5-6 times [2].

This promoter can be utilized by other iGEM teams as a standardized promoter sequence for constitutive gene expression in C. reinhardtii.

Reference List:

1. Schroda, M., Beck, C. F., & Vallon, O. (2002). Sequence elements within an HSP70 promoter counteract transcriptional transgene silencing in Chlamydomonas. The Plant Journal, 31(4), 445-455.

2. Lumbreras, V., Stevens, D. R., & Purton, S. (1998). Efficient foreign gene expression in Chlamydomonas reinhardtii mediated by an endogenous intron. The Plant Journal, 14(4), 441-447.

Sequence and Features