Team:SZU-China/Parts

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Basic Parts

Eight Basic parts has been built this year.

1. Basic Parts for light-mediate controlled yeast-two-hybrid system.

• ①BD-CIB1 (BBa_K1884001)
• ② AD-CRY2(BBa_K1884002)
• ③ Upstream activating sequence(BBa_K1884004)

Two fusion proteins and a promoter which is activiated in special situation are used to built our light-mediate controlled system.

2. Basic Parts for regulating genes in photosynthesis

• ④PsbA(BBa_K1884005)
• ⑤Ferredoxin-NADP(+)Reductase (BBa_K1884006)

Two coding genes transcrpting into two microRNA which would targeting two gene in photosynthesis coding D1 protein and FNR enzyme.

3. Basic Parts for regulating genes in photosynthesis

• ⑥ Paromoycin(BBa_K1884013)
• ⑦ Hygromycin B(BBa_K1884003)

Two antibiotics coding genes is used as a reporter and resistance genes to select bacteria and green algae.

4.Basic Parts for New promoter in Chlamydomonas

• ⑧RBCS2 promoter(BBa_K1884010)

A promoter is for use in activiating our antibiotics coding gene.

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Composite Parts

Five composite parts has been built this year.

1. Two devices to produce hydrogen

• ①PSAD Promoter+BD-CIB1+PSAD Terminator(BBa_K1884007)
• ②PSAD Promoter+AD-CRY2+UAS+PsbA+PSAD Terminator(BBa_K1884008)
• ③ PSAD Promoter+BD-CIB1 +PSAD Terminator(BBa_K1884008)
• ④PSAD Promoter+AD-CRY2+UAS+FNR+PSAD Terminator(BBa_K1884009)

For activating the light-mediate controlled system, followed the example of yeast two-hybrid system, Two plasmids which contain two fusion protein coding gene have built in green algae.the microRNA gene is constructed on the 3 primer end of UAS, When the system is activated, transcription is on and product microRNA in order to regulate two pivotal genes in photosynthesis.

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2. Two devices for use in produce antibiotics

• ①RBCS2 Promoter+Paromoycin+RBCS2 Terminator(BBa_K1884011)
• ②RBCS2 Promoter+Hygromycin B+RBCS2 Terminator(BBa_K18840012)

In order to detect the function of our system, we construct these two devices into two different plasmids. If we select the right colony in the TAP medium plate contain Paromoycin resistance and Hygromycin B resistance.

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Favorite	Name	Type	Description	Design
√	BBa_K1884007	Composite	PSAD Promoter+BD-CIB1+PSAD Terminator	Yongjing Ping
√	BBBa_K1884008	Composite	PSAD Promoter+AD-CRY2+UAS+PsbA+PSAD Terminator	Changxing Hu
√	BBa_K1884009	Compostie	PSAD Promoter+AD-CRY2+UAS+FNR+PSAD Terminator	Changxing Hu
	BBa_K1884001	Coding	BD-CIB1	Chengrong Xie
	BBa_K1884002	Coding	AD-CRY2	Chengrong Xie
	BBa_K1884004	Coding	Upstream activating sequence	Chengrong Xie
	BBa_K1884005	Coding	PsbA	Changxing Hu
	BBa_K1884006	Coding	Ferredoxin-NADP(+)Reductase	Changxing Hu
	BBa_K1884013	Coding	Paromoycin	Yongjing Ping
	BBa_K1884003	Coding	Hygromycin B	Yongjing Ping
	BBa_K1884010	Coding	RBCS2 promoter	Yongjing Ping

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Part Collection

Followed the example of yeast two-hybrid system, 2016 SZU-China creates a part collection named light-mediated expression system collection, including two fusion proteins, and the Upstream activating sequence.

• BBa_K1884001;  BBa_K1884002;  BBa_K1884004;

These are the basic parts numbers of fusion proteins and light controlled proteins. Fusion proteins (BBa_K1884001; BBa_K1884002) are based on yeast-two-hybrid interaction for use in controlling DNA transcription by blue light. BBa_K1884004 is consist of a BD binding site and a promoter to start the transcription which can bind with the Activity domain .

• BBa_K1884007;  BBa_K1884008;  BBa_K1884009;

Based on the light-mediated expression system, we built 2 devices  (BBa_K1884007 and BBa_K1884008; BBa_K1884007 and BBa_K1884009) which have two different microRNA for hydrogen production in order to control the production of hydrogen.

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