To my mind, the most appropriate way of addressing this research is to isolate fractions (aqueous, alcohol, etc.) and test these fractions for anticancer activity in a model system. This will help narrow down the type of compounds that may be responsible for anticancer activity of a particular plant you are interested in. Select the fraction that will give you the highest activity. Subsequently, you may try purifying the compounds from that active fraction. Different compounds from the most active fraction may again be subjected to anticancer activity, again using the model system. This may help you reach closer to the active principle. This is a time consuming approach but will be more directional and acceptable. Chemical characterization of the active principle requires well-equipped lab and sophisticated instruments but that type of services can be hired also in case you do not have those equipment. Once the compound is identified and characterized, then the biotechnological approach will be quite useful to enhance the level of of compound through over-expression of genes involved in the biosynthetic pathway of the active principle.

This a 2-4 years project (or may be more) but do not loose heart. Your passionate approach can certainly give you wonderful results. All the very best in your endevour.

The anticancerous effect will be probably rather because of some low molecular substance the because of gene. But you can try several ways to extract either proteins or low-weight substances and test their activity against cancer.

Well you can work at the chemical and biochemical level , or testing metabolic extracts from plant checking which part of the plant might have activity and as well following the same procedure with protein extracts after purification and testing protein activity according with the anticancer test you are willing to do .

Yes dear , what r the procedure to find protein purification, and what r the biochemical test we can do for this

Dear Karen, I am working in this area and as you mentioned you work on chines medicin and I generally can assumed that this may be related to secondary metabolities in perticulary will be to glycosides and i generally can say its related to saponin metabolites (and for this you have to check phytosterol pathway) and this is quite large pathway but you can make some choice if you check the data base of NCBI

It doesn't make any sense to look for a gene, as posted before the anti cancer effect must be a chemical compound produced by the plant, so you have to look for. I don't think a protein is the agent of the anti cancer effect because proteins are degraded during digestion, so a chemical will more likely to be responsable. The problem is you have thousands of compounds produced by plants, in some case under specific, and often unknown, conditions. To go further read the following articles:

Rodríguez, A. & Infante, D. (2009). Network models in the study of metabolism. Electron. J. Biotechnol. 12(4):1-19.

Rodríguez, A. & Infante, D. (2011). Characterization in silico of flavonoids biosynthesis in Theobroma cacao L. Network Biology 1(1):34-45.

Both are free on the net.

well Diogenes Infante , but this all r microbial based we want to do on plant , it will work , and how to do this all thing in plant cell, sorry i am very new for this all .

Dear Mostafa Rahman , can i get some protocol for this all .

Look for a teacher at your university

Dear Karen,

This is not a trivial task. It would certainly help to know what is causing the anti-carcinogenic (if that is what you mean?) effect. It could be a protein, a metabolite, or perhaps even a combination of these? Once you know the causal component, you could try to see if anything is known about the biochemical or genetic pathway it is involved in, to get an idea of what kind of genes could be involved, but this may take a long time. Of course you could gamble and use a shotgun approach in which you make an expression cDNA library in a suitable (eukaryotic?) host and test extracts of the individual expressors (expressing one cDNA) for their effect on a proper tester (e.g. human cancer cell lines). This could work if the gene will do the same thing in the host as it does in plants. When secondary metabolites are the causal agent, this is unlikely to work. If it is a protein which does the job, it might work.

Then there is also the genetic way, in which you cross the medicinal plant with a related plant not showing the medicinal effect. Provided this is possible, you could create a genetically segregating population which you could screen with genetic markers (to be made if this is an unexploited species, but current low costs for whole genome sequencing approaches may be useful) to find associations between markers and the medicinal effect. That would identify markers in the vicinity of the genes causal for or contributing to the medicinal effect. If your markers are genetically very close to the causal genes you could use them to probe a genomic DNA library of your medicinal plant to eventually cover the region containing the causal gene(s) with overlapping DNA clones. You can sequence these to identify the genes and use these to query their function. But, again, not a trivial task...

To my mind, the most appropriate way of addressing this research is to isolate fractions (aqueous, alcohol, etc.) and test these fractions for anticancer activity in a model system. This will help narrow down the type of compounds that may be responsible for anticancer activity of a particular plant you are interested in. Select the fraction that will give you the highest activity. Subsequently, you may try purifying the compounds from that active fraction. Different compounds from the most active fraction may again be subjected to anticancer activity, again using the model system. This may help you reach closer to the active principle. This is a time consuming approach but will be more directional and acceptable. Chemical characterization of the active principle requires well-equipped lab and sophisticated instruments but that type of services can be hired also in case you do not have those equipment. Once the compound is identified and characterized, then the biotechnological approach will be quite useful to enhance the level of of compound through over-expression of genes involved in the biosynthetic pathway of the active principle.

This a 2-4 years project (or may be more) but do not loose heart. Your passionate approach can certainly give you wonderful results. All the very best in your endevour.

do you mean the gene that involve in the biosynthesis of metabolites against cancer? if so, what is the potent chinise medicine in cancer treatments? polyhydroxy aklaoids are among the most promissing anti-cancer metabolites that currently used such as the N-methyl-hydroxynojirimycin ... if that is the case, you must trace back the enzyme related to the pathway, the gene that involve inthe biosynthesis of the intermediate may be manipulated and tranferred into other cells such as yeast for metabolites production, but all secondary metabolites are from primary metabolites...

This is a formidable quest. Big pharmas pour billions of dollars on this "holy graal"... but that doesn't mean one should no be intimidated... However, the sole objective of big pharmas is profit, not canker cure, by selling expensive (usually) medicines. So, there are chances...

I dont know whether it will work for you or not! but i have some suggestion regarding your work. Usually the medicinal plant contains different types of secondary metabolite which are responsible behind the anti-cancer, anti-microbial effect and so on. So I think First, you should to find some literature on medicinal plant specially which focused on anti-cancerous activity. it will aid you to make idea about the chemicals which are responsible for anti-cancer activity. then you have to find the pathway through they are synthesizing, as well as the gene responsible for their expression. thus you can identify the gene/genes and use RT PCR to check whether they are expressing. if you need further help, im available on [email protected]. hope together we can make a way, if you contact with me... hope for the best

@Diogenes Infante.. thanks mam for ur suggesttion 2 go for first 2 scrreen out metabolic pool for finding better anticacncerrous copmpound nd thn 2 go for anticancerous gene

@Mark Aarts... really liking ur suggestion for creating shotgun approach nd then go for creating cDNA librarary and checking association with marker nd many thanks for it

@Pankaj Vaishnav .. its very informative wid ur suggestion tht how 2 narrow down metabolic pool nd thn carry out chemical charactrization for its better characterization nd without its work would be more confusing and also thnks for ur suggstion for carry out over exprssion of targeted gene nd definitly sir keep faith me nd i will not loose my heart in my work

@Aziz Ahmad ... its quite impressive to solve problem with traccing back enzyme of pathway from KEGG or othr database but which is more confusing is many variant form of enzyme nd its diffrent conditional expression to solve for it nd if u suggest me over this i m highly obliged to u

@Munvar Shaik .. its nthing like persnal dear nd i will defintly go for checking pharmacokinetic nd toxicity of extracct nd wid identification of anticancer gene thn want 2 go for creating its variant library nd going for checking its expression wid microarray possibly if fortunate

@Mostafa Rahman ...thnks for ur suggesstion 2 explore NCBI database but as u are working on same problem may be u can give me some good suggestion over specific metabolite daatabase if u hav used

@Tariqul Tareq ...thank u for valuable suggestion for explioring nd cheking gene expression with RTPCR nd thnks for giving ur email id nd defintly i will ask u for ur help

thnks 2 everyone for ur time nd ur valuable suggestion

You want to identify the anti-cancerous gene from the plant..that means you want to identify anticancer drug synthesis gene. better you would go with highly proved plant for example catharanthus roseus for the identification drug synthesis gene.You can identify Vinblastine, Vincristine, Podophyllotoxin, Taxol, Camptothecin etc. synthesis gene. you should study that what are the plant synthesis anticancer drug available..what are the genes involving synthesis of the same..then you will go with your way..good luck

@Raju Karthic ..it is very useful for me to use comaprative approach wid ur suggestion nd thnks for ur valuable suggestion of list of secondary metabolite with anticancerous activity but if u have any idaea for identified gene responsible for their activty

thanking u for ur time

I kindly ask everybody to use plain english

Ph.D is a degree and a licence to do research and teaching. You are in search of subject to do phd. better you find your interest of subject and do your phd. this will be helpful to achieve your target. do not postpone your phd in search of subject. For a achievement subject is not a issue. your interest and hard work is a main key factor. you came for consultation is nice..my suggestion is applied research in your interested subject. come with your interest it will be more helpful to you..please do not divert karen's topic..you can start new..good luck..

Dear Sir,

We are working in our lab on systematics and diversity of plants, using molecular markers we try to resolve the taxonomic and systemacs problems among various taxa. Your problems is like a gene expression problem. So you must approach a molecular biologist working on gene expression.

Baleshwar

@Baleshwar Meena....respected sir, so many thanks forr ur sugestion to chk gene expression wid molecular biologist, can u suggest me sm good mol. biologist at ur lab, nd pls. i m just phd fellow so dnt call me sir

thank u...

This is good theme for research based on high throughput molecular biology, gene expression, transgenics and system biology. An integrated approch in a network mode project with huge funding is required for this job.

@Joanna Cross...with so many previous research on cellline with liver cell it already proved to be anticancerous nd showing good activity toinhibit cancer cellline growth and invasion so i choice 2 wrk on it...

thank u

@Baleshwar Meena... sir, we have already planned for it and fund is coming as DST secctioned project so no woory of it nd its better to work on integrated project or not?

thanking you

With tissue culture we can acount and observer the many kind of cancerous cell

There is nothing like a anticancer gene in plants, at the most one or few genes may be involved in producing the anticancer compound in the plant, first try to purify the compound, identify the compound, then think of isolation of gene /genes involved in its pathway. Goodluck.

Dear, Dahlia,

I could not understand your question, will you be more specific?

As previously mentioned what is the meaning of anticancer gene?? you have anticancer compounds and these compounds could be diverse starting from primary end with secondary metabolites? so you have to specificy which group ofcompound is interested in your material through fractionation and MTT assay then you can look for the gene which is synthesis these compounds whic inmost of cases will be multiple genes interfer.