Table 1 Synthetic oligonucleotides used in this study.

From: The novel transcriptional factor HP1BP3 negatively regulates Hsp70 transcription in Crassostrea hongkongensis

Primer

Sequence 5′ → 3′

Position(s)

Purpose

Oligo(dT)18

TTTTTTTTTTTTTTTTT

 

Reverse transcription

P70-promoter up

TTCCATGCACTCTAGCTAAG

−304–285

Amplification of ChHsp70 promoter fragment for DNA-affinity purification and EMSA

P70-promoter down

GAATCGTAGTCTAGTTGACTC

+40–+60

HP1BP3-up

ACATATTTGAATGTATGTATGAAT

−14–+10

Amplification of ChHp1bp3 coding sequence

HP1BP3-down

ACTGTTGTGGATAAAGATTCCCT

+1528–+1550

UP

CGGCAGTGGTATCAACGCAGAGTAC

 

Universal primer used with 3GSP2 and 5GSP2

UP(dG)

CGGCAGTGGTATCAACGCAGAGTAC(G)10

 

5′ RACE PCR

5GSP1

TATTTCTTCATCATTAGCA

+594–+612

5GSP2

TTCCCATGTTTTTCCGCCTCTGAC

+789–+812

UP (dT)

CGGCAGTGGTATCAACGCAGAGTAC(T)18

 

3′ RACE PCR

3GSP1

TACATAGAAAAGTATTAC

+898–+915

3GSP2

TCAGATGACATGTGGCAGGCTGTT

+967–+990

HP1BP3-F-NdeI

GGGAATTCCATATGTATGAATAT

+1–+12

Amplification of ChHp1bp3 for protein expression

HP1BP3-R-XhoI

CCGCTCGAGTTTTCTGGAGAC

+1420–+1431

HP1BP3-E-KpnI

ATAAAAGGTACCATGTATGAATATTACACAGAATCCATC

+1–+27

Amplification of ChHp1bp3 for overexpression

HP1BP3-E-XhoI

ATAAAACTCGAGCTATTTTCTGGAGACTGGGGC

+1414–+1434

Actin-Up

CTGTGCTACGTTGCCCTGGACTT

+701–+723

RT-PCR

Actin-Down

TGGGCACCTGAATCGCTCGTT

+829–+849

RT-HP1BP3-Up

AAACAGCCTCAGGACAGATG

+1094–+1113

RT-PCR

RT-HP1BP3-Down

CAAGTTGAAATGATCCGCCAC

+1238–+1258

RT-HSP70-Up

GCCAAACTACATCAGAACGGGTC

−1860–1838

RT-PCR

RT-HSP70-Down

TCCATCTCCTCTACAGTCGGTCC

−2461–2439