Promoters and usage thereof

Abstract

Promoters and usage thereof. Two promoters, acuF and hsp promoters, comprise the nucleotide sequences of SEQ ID NO: 1 and 2, respectively.

Description

technical field

[0001] The present invention relates to promoters and their use. Background technique

[0002] In Asian countries, Monascus has long been used as a food additive. The grain fermented by Monascus is bright red to purple, and retains the original shape of rice. In addition, Monascus ferment products are also believed to have health-promoting properties. The application of Monascus in rice wine production is due to its high amount of α-amylase, which can promote the conversion of starch into glucose. Current studies have confirmed the medical effects of Monascus fermented products such as Monacolin K. Moreover, the preservation effect of the fermented product of Monascus has also been proved by experiments.

[0003] The main research focus in this field is the gene product or cultivation and production methods of Monascus, however, the research on Monascus sequence tag (EST) gene bank is in the ascendant, and the sequence tag of Monascus may provide more inf...

Examples

Embodiment 1

[0059] Example 1: acuF promoter

[0060] 1. Materials

[0061] (1) Host: Monascus pilosus BCRC 38072, Monascus pilosus, BCRC 31527, Neurospora crassa BCRC 32685 (Food Industry Development Research Institute).

[0062] (2) Competent cells: ECOS TM E.coli competent cells DH5α (Yisheng Biotechnology Company).

[0063] (3) Plasmid: pHygEGFP (BD Biosciences), pGEM _ -T EasyVector(PROMEGA).

[0064] (4) Medium:

[0065] a. Culture medium for Escherichia coli: LB broth (USB), agar (USB).

[0066] b. The medium of Monascus and pink bread mold (Neurospora crassa): PDA (DIFICO), PDB (DIFICO), Vogel's medium (see * ), and topagar.

[0067] (5) Antibiotics: penicillin (Ampicillin), hygromycin (Hygromycin B) (SIGMA).

[0068] * Vogel's Medium

[0069] 1. Trace element solution: 5g citric acid·H 2 O, 5g ZnSO 4 ·7H 2 O, 1gFe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 Q. 250mg CuSO 4 ·5H 2 O, 50mgMnSO 4 ·H 2 O, 50 mg H 3 BO 3 , 50mg Na 2 MoO 4 2H 2 O was dissolved in 95ml of sterile...

Embodiment 2

[0143] Example 2: Hsp promoter

[0144] 1. Materials:

[0145](1) Host: Monascus BCRC 38072, Monascus pilosus BCRC 31527, and Neurospora crassa BCRC 32685 (Food Industry Development Research Institute).

[0146] (2) Competent cells: ECOS TM E.coli competent cells DH5α (Yisheng Biotechnology Company).

[0147] (3) Plasmid: pHygEGFP (BD Biosciences), pGEM _ -T EasyVector(PROMEGA).

[0148] (4) Medium:

[0149] a. Culture medium for Escherichia coli: LB broth (USB), agar (USB).

[0150] b. The medium of Monascus and pink bread mold (Neurospora crassa): PDA (DIFICO), PDB (DIFICO), vogel's medium (see * ), and topagar.

[0151] (5) Antibiotics: penicillin (Ampicillin), hygromycin (Hygromycin B) (SIGMA).

[0152] * Vogel's Medium

[0153] 1. Trace element solution: 5g citric acid·H 2 O, 5g ZnSO 4 ·7H 2 O, 1g Fe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 O, 250mg CuSO 4 ·5H 2 O, 50mgMnSO 4 ·H 2 O, 50 mg H 3 BO 3 , 50mg Na 2 MoO 4 2H 2 O was dissolved in 95ml of sterile deio...

technical field

[0001] The present invention relates to promoters and their use. Background technique

[0002] In Asian countries, Monascus has long been used as a food additive. The grain fermented by Monascus is bright red to purple, and retains the original shape of rice. In addition, Monascus ferment products are also believed to have health-promoting properties. The application of Monascus in rice wine production is due to its high amount of α-amylase, which can promote the conversion of starch into glucose. Current studies have confirmed the medical effects of Monascus fermented products such as Monacolin K. Moreover, the preservation effect of the fermented product of Monascus has also been proved by experiments.

[0003] The main research focus in this field is the gene product or cultivation and production methods of Monascus, however, the research on Monascus sequence tag (EST) gene bank is in the ascendant, and the sequence tag of Monascus may provide more inf...

Examples

Embodiment 1

[0059] Example 1: acuF promoter

[0060] 1. Materials

[0061] (1) Host: Monascus pilosus BCRC 38072, Monascus pilosus, BCRC 31527, Neurospora crassa BCRC 32685 (Food Industry Development Research Institute).

[0062] (2) Competent cells: ECOS TM E.coli competent cells DH5α (Yisheng Biotechnology Company).

[0063] (3) Plasmid: pHygEGFP (BD Biosciences), pGEM _ -T EasyVector(PROMEGA).

[0064] (4) Medium:

[0065] a. Culture medium for Escherichia coli: LB broth (USB), agar (USB).

[0066] b. The medium of Monascus and pink bread mold (Neurospora crassa): PDA (DIFICO), PDB (DIFICO), Vogel's medium (see * ), and topagar.

[0067] (5) Antibiotics: penicillin (Ampicillin), hygromycin (Hygromycin B) (SIGMA).

[0068] * Vogel's Medium

[0069] 1. Trace element solution: 5g citric acid·H 2 O, 5g ZnSO 4 ·7H 2 O, 1gFe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 Q. 250mg CuSO 4 ·5H 2 O, 50mgMnSO 4 ·H 2 O, 50 mg H 3 BO 3 , 50mg Na 2 MoO 4 2H 2 O was dissolved in 95ml of sterile...

Embodiment 2

[0143] Example 2: Hsp promoter

[0144] 1. Materials:

[0145](1) Host: Monascus BCRC 38072, Monascus pilosus BCRC 31527, and Neurospora crassa BCRC 32685 (Food Industry Development Research Institute).

[0146] (2) Competent cells: ECOS TM E.coli competent cells DH5α (Yisheng Biotechnology Company).

[0147] (3) Plasmid: pHygEGFP (BD Biosciences), pGEM _ -T EasyVector(PROMEGA).

[0148] (4) Medium:

[0149] a. Culture medium for Escherichia coli: LB broth (USB), agar (USB).

[0150] b. The medium of Monascus and pink bread mold (Neurospora crassa): PDA (DIFICO), PDB (DIFICO), vogel's medium (see * ), and topagar.

[0151] (5) Antibiotics: penicillin (Ampicillin), hygromycin (Hygromycin B) (SIGMA).

[0152] * Vogel's Medium

[0153] 1. Trace element solution: 5g citric acid·H 2 O, 5g ZnSO 4 ·7H 2 O, 1g Fe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 O, 250mg CuSO 4 ·5H 2 O, 50mgMnSO 4 ·H 2 O, 50 mg H 3 BO 3 , 50mg Na 2 MoO 4 2H 2 O was dissolved in 95ml of sterile deio...

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