The USPTO Registered Logo Badge of the Medical Marijuana Initiative of North America - International Limited, an Arizona Benefit Corporation to the left of Mt Hood sunlit from the northwest in June followed by the proprietary MMINAIL octocopter delivery bot.

Target Nucleus: Using A. tumefaciens as a Vector for Introducing the Over-expression of Limonene in Cannabis.

Welcome! This is the Target Nucleus page of the Carbon Free Footprint project.

The total area of cultivated land in the year #2004 was 1,400 million hectares ( 1.4 billion hectares ).

Surely the world's scientists will find a way to muscle in a little Cannabis plantation here or there throughout the globe in our current millennia.

More to come ...

A. tumefaciens

A. tumefaciens is a common bacteria that causes crown gall disease.

Dicots are especially susceptible, both plants and trees, though Monocots may be infected, as well.

Dicots are 'di-cotyledonous' woody shrubs and trees that have spread across the globe.

Dicots have broad leaves with branching veins like an Apricot tree.

The serrated leaves and branching veins of a Cannabis plant, though an annual herb like the Basil plant, also qualify for inclusion in the family of Dicots.

On the other hand, narrow leaved plants with parallel grains such as grasses, grains and cedar trees are known as Monocots.

Monocots are 'mono-cotyledonous' plants that have spread across the globe, as well.

The Magic of Bacteria

A. tumefaciens does its magic by way of a 'Tumor Inducing', or TI plasmid.

A plasmid is a circle of DNA that contains genes, yet is separate from the parental chromosomes of the bacteria and can, therefore, be replicated on its own.

The size of the TI plasmid is between 200 and 800 kb ( base pairs ).

However, to perform the infection, a region of between 12 and 24 kb ( base pairs ) is used.

This 'Transfer DNA' region, or T-DNA is integrated directly into the nucleus of the host plant during the infection process.

The goal of the integration is to change the parental chromosomes of the host that are present in every nucleus, of every cell of the host organism.

But, here, specifically, the cells of the growing organism in the crown area between the trunk and the root system are targeted by the bacteria.

One function of the genes that are being integrated into the host genome is to form the visible gall structure.

A second function of the genes that are being integrated into the host genome is to synthesize opines.

Opines are modified Amino Acids that manufacture both carbon and nitrogen molecules that can now feed the expanding colony of A. tumefaciens.

The host plant now spends its ATP energy packs creating a food for the bacteria, in a form of which the host plant itself cannot use.

This is a naturally occurring state of hijacking of the bio-chemical machinery of the host plant by the bacteria!

Outside the transfer region, on the polar opposite side of the plasmid ring, are genes that act upon the process of opine catabolism.

This assisting 'Virulence Region' of DNA and its compliment of genes sits on the plasmid ring, as well, but far away from the T-DNA transfer region.

Note. Opine catabolism can be enhanced through the manufacture of enzymes that break down the opine.

Pretty clever, heh? The bacteria retains the genes for manufacture of the enzymes responsible for breaking down the opine away from the integrating transfer region of the plasmid.

Therefore, the host plant is not able to break down the copious amounts of opine that the tree is now genetically forced into making!

Also outside the transfer region of the plasmid, are the genes responsible for encoding Virulent proteins ( Vir-p ) that facilitate the transfer of the T-DNA snippet through the transport channel.

Thus, the integration apparatus assisted by the the Vir-p is able to transfer the TI region of the plasmid into the host plant via the transport channel outside the purview and control of the host plant, as well!

Gotcha! Punch that plant cell wall and membrane!

Target: Nucleus!

Hitching a Ride

Two can play this game of hijacking the genome!

Why not isolate the gene responsible for kicking in the manufacture of Limonene, an un-foreign substance naturally resident in the Cannabis plant, to the point of Over-expression?

While inserting the gene for the Over-expression of Limonene, why not add a marker that will eliminate all copies of the target genome not successfully integrated?

This can be achieved by way of another, simultaneous gene insertion.

Specifically, the gene that induces resistance to the anti-biotic ( bacteria killer ) Kanamycin.

Kanamycin is also toxic to plant cells.

By incorporating Kanamycin-resistance into the genome of the host plant, those plant cells not affected by the transfer plasmid will wither.

Upon application of a dose of Kanamycin, introduced genes that manufacture enzymes that can in effect 'disarm' the killer Kanamycin will move into play.

Thus saving your preferred callus from certain death!

Any explant not containing the genes for Kanamycin-resistance, and by extension the genes for Over-expressing Limonene, will be destroyed by the application of Kanamycin.

Pretty final, heh?

Think of that the next time your daughter decides to date a plant biologist.


The Knockout

Why not 'knockout' the tumor producing genes, the virulence region, and the genes coding for both opine synthesis and opine catabolism from the plasmid prior to integration with the host plant?

Good idea!

Let's do just that!

By 'knocking out' the tumor producing genes, the virulence region, and the genes coding for both opine synthesis and opine catabolism we have created a benign plasmid.

One that can be loaded with the genes to Over-express Limonene, in addition to the identifying Kanamycin-resistance genes.

Simply place the genes into the transfer region ( T-DNA) as described.

Don't forget the Vir-p's!

Remember, the Virulent proteins, or Vir-p's assist the transfer of the entire plasmid by facilitating movement of the plasmid through the transfer channel between bacteria and host.

Toss a few of those into the balance of the plasmid for good measure.

By conjugating a common E.coli bacteria containing the target plasmid with a knocked out A.tumefaciens bacteria containing only a second 'Helper Vector' ...

One that is full of the genes required to replicate Vir-p's, the now combined A.tumefaciens is ready for injection into the host plant.

Note. The genes required to produce the Vir-p's will be activated upon insertion.

The virtual goal of the balance of this report will be to reproduce a single Transgenic Cannabis Plant, or TCP phenotype capable of producing Limonene.

Next, a True-breeding line of Cannabis plants that truly Over-express Limonene via the introduction of a promoter sequence into the genotype of the line will commence.

Finally, a synthetic version of the Over-expression gene containing DNA triplets more common to plants will be introduced to the line resulting in a 100-to-1,000 times increase in the manufactured expression of Limonene by the plant.

The true-breeding line of TCPs can then be propagated from the lab via Explant Clones, or seeded from fresh achenes after further back and cross-breeding.

The extraction of Limonene and other terpenes and cannabinoids from the herbaceous plant material of the Cannabis plant by mechanical methods utilizing CO2 under pressure are now a reality in today's modern laboratories.

Scaling up our processes to accommodate a growing market for pure Limonene and other novel terpenes and cannabinoids is the wave of the future.

More to come ...

Supporting Content Small The Flammarion Logo Badge Small The Flammarion Logo Badge

Welcome! This is the Supporting Content section for the Target Nucleus Page | Carbon Free Footprint Project page of the MMINAIL.

The Concept Library of the MMINAIL is a static informational website.

As such, a Supporting Content section is placed below each (.html) page in the flow of the default layout to grant the end-user additional pertinent information when navigating this site.

E Books

With today's explosion of modern devices ...

Some consumers like to read their articles on their Kindles or iPads.

Therefore, a method to convert any page of this website into an eBook ...

Is auto-built into the top of the Navigation <aside> ...

As well as in the <footer> of every page.

Dot ePub

Simply click the following eBook icon = small dot epub button eBook icon to render the current page as an Overdrive (.epub) or a FB Reader (.mobi) .

Note. The Apple iBooks platform also reads and displays the (.epub) format, as well.

Secure Hyperlinks

The A's Have It!

In today's complicated world of coding, even the use of the standard hyperlink ...

<a href="" title="" target=""></a>

Is placed into question by the standards of newer, more modern browsers such as Google's V8 powered Chrome browser.

Safe Internal Links

Therefore, to make life easy for the end-user ...

The MMINAIL will show safe, internal links in the bootstrap default color of light dodger blue.

Safe External Links

In addition, safe external links ...

When designated https secure ie.) https://.

Will also be shown in the color of light dodger blue.

Standard External Links

All other external links ...

If designated with standard http ie.) http:// ...

Will be shown in the color of indian red.

How To Navigate This Website

At this website ...

We have the <html> hub for the Benefit Corp aka our Concepts Library.

You (Usted), as a current visitor to our Concepts Library, are now at the Target Nucleus Page | Carbon Free Footprint Project page.

Navigation Aside

In the Navigation <aside> section of this website ...  ...

The end-user will find a Stack of Topics.

Or, for you smartphone users ... the Navigation <aside> section is up! ...  

There, the smartphone user will find the same Stack of Topics for the Concepts Library at MMINAIL.

Stack O Topics

The Navigation <aside> section to your right ... ... If you are viewing this page from the traditional landscape style of your laptop.

Or, the Navigation <aside> section from above ... ... If you are viewing this page from the modern portrait style of your smartphone ...

Both house the same Stack of Topics available for selection at the Concepts Library of the MMINAIL.

Instruction Set

To access one of the Articles or Lessons available for your preferred Topic of Interest ...

  • Simply scroll through the tabs of Topics above, or to the side ...

  • And, click on one that you may have an interest in.

Back Home

Note. At each Topic of Interest, the end-user will find a hyperlink back to the Home page of the Concepts Library.

By clicking on the Home icon   the end-user will be returned to the Home page of the Concepts Library.


When an end-user clicks on a Topic of Interest ...

Either above  ...

Or, over at  ...

The Stack of Topics in the Navigation <aside> section ...

The menu explodes vertically to reveal several subtopical choices.

To select any other Topic of Interest, or any of the many subtitles ...

  • Simply click on the corresponding glyphicon to open up the referenced file.


If you get stuck ...

  • Simply click on the Home icon   to be hyperlinked back to the Home page of the Concepts Library.

Got An Idea?

If you have an idea that will spruce this site up even better than it already is ...

Then, you know the drill ...

Simply go ahead and fork this repo, make your changes, and send us a pull request.

25 Mile Rule

"When residing 25 miles extant of a state legalized dispensary ..."

Spanning the entire connectome of the modern Medical Marijuana Initiative in a connective cartography of ideas and hyperlinks ...

The open source authors of this project hope to enlighten the world to the plight of the common medical patient.


Due to a labyrinth of government regulations and costs ...

Today's common medical patient is virtually devoid of healing phyto-cannabinoids and other beneficial terpenes globally for no other reason than his or her current residential address.

Doesn't that sound a bit strange to you?

Restricting Medicines

Restricting access to medicine ...

Almost Un-constitutional, isn't it?

And, with patients in Arizona paying anywhere from $75 to $90 USD for an eighth (1/8th) of an ounce of medicine ...

Don't you think it is time that we allow ALL patients the right to grow, stash, and cure their own medicines?

Make Contact

The MMINAIL is dedicated to the preservation of the hard-earned rights of Medical patients.

Don't you think it is time that we allow ALL patients the right to grow, stash, and cure their own medicines?

We do at the MMINAIL.

Patient Rights

The Authority To Cultivate ...

"The right of a patient to grow up to twelve (12) state-legal Medicinal plants for their continued health and growth shall not be abridged."

When the voters of Arizona by the slimmest of majorities chose to grant Medicinal access to qualified patients throughout the fifteen (15) counties of Arizona ...

No one expected the Arizona Health and Human Services Department to abscond with both the patients' then given Authority To Cultivate as well as overcharging for the costs to operate the Medicinal "green" card program.

But, they did ... on both accounts.


If you agree, then why don't you donate a little spare cash for the cause?

Or, a little of your coding time?

Or, simply contact your favorite Arizona State Senators and Congressional Representatives today!

Coding Community

If you are an expert in your field of specialty ...

Why not fork our Concept Library and create a new Topic of your choosing, or embellish upon an existing Topic.

Pull Requests

Send us a Pull Request.

Our team of coders will review your contribution and get back to you with a "thumbs up" ( or, "thumbs down" ).

Can You Contribute Code?

As we can now decipher the time spent by our functions at the code block level ...

How much of your coding time are you willing to invest in this project?

As we rely heavily upon contributions from the coding community, your contribution of code will be greatly appreciated.

Thank you for your effort and participation in our program to code the globe!

Can You Contribute Cash?

If you have enjoyed and benefited from this article ...

Then, why not drop a little change in our digital bucket at the Square Up - Cash Me donation portal.

As we can now spend the equivalent of our grandparents' monthly mortgage payment at our local Sunday football games ...

How much of your spare cash are you willing to invest in this project?

Starving Coders

As we rely heavily upon contributions from the coding community, your contribution of cash will be greatly appreciated.

Note. Your donation will help us fund our Starving Coders initiative through our Medcoin™ Crypto Currency Division.

Can You Contribute Ether?

As we have witnessed the exponential growth of Bitcoin (BTC) from 9 cents USD per coin ...

To over $18,000 USD per coin over the past nine (9) years.

Plus, the recent explosion of Ripple (XRP) from less than one-penny to over $2 USD.

How much of your spare Ether ( ETH ) are you willing to invest in this project?

As we rely heavily upon contributions from the coding community, your contribution of ether will be greatly appreciated, as well.

Public Hash

Simply scan the following QR Code to extract our Coinbase public hash address to send Gifts O Wei in support of our Medcoin™ Crypto Currency Project.

QR Code - Coinbase - Bus Card

No Warranty

Disclaimer. The author of this website has made every effort to ensure accuracy ...

However, the information contained in this website, and in its pages, is offered to the public without any warranty expressed or implied.

Therefore, the author of this website, and by extension its pages and content cannot be held liable for any damages that may be caused indirectly or directly by the software instructions or tutorials contained in the pages of this website.


The license and privacy policy pages ...

For further review, please see the License page and the Privacy Policy page of this website to clarify.

Screen Responsiveness

The pages of the Concept Library have been tested for mobile responsiveness.

The authors have determined the accuracies of the page renderings at both a portrait width of 360px and a landscape width of 640px.


Testing devices have included selections from the following list of devices ...

  • Amazon Kindle Fire HDX 7, 3rd Generation tablet

  • Virtual Galaxy S5 smartphone emulator

Kindle Tablets

On the Kindle tablet, both portrait and landscape renderings appear accurate.

Amazon Fire Phones

And, on the Amazon Fire phone, using automatic screen rotation, both portrait and landscape renderings appear accurate.

Foreign Language

The foreign language expression ¡Finito! can hold a translation from Spanish-To-English ...

Via the <title> tag.

Simply hover over the word in Spanish to reveal its English equivalent.

For example ...

Hovering over the Spanish phrase ¡Por ejemplo! reveals the English equivalent of "For Example".

Try it!


For an acronym to be visually effective when reading a line of text ...

The acronym must first be declared.

In addition, the acronym must stand out from the body of information.

To accomplish both of these objectives, the MMINAIL has selected Ashley Gold.

Ashley Gold

Ashley gold is a primary color within the registered Logo Badge of the Benefit corporation.

Whenever the end user spies a designated Acronym of Ashley Gold ...

Simply hover over the Acronym to reveal the underlying meaning embedded in the title element of the <abbr> tag.


The <abbr> tag can also be used to house Definitions as well as Acronyms.

As we have seen in the above referenced Ashley Gold definition ...

Whenever the end-user spies a designated Acronym or Definition of Ashley Gold color ...

Simply hover over the Acronym or Definition to reveal the underlying meaning embedded in the title element of the <abbr> tag.

Carriage Return

The carriage return, or ... ➡️

The carriage return is used extensively throughout the working repos of the Concept Library to prevent the overflow of raw code.

By preventing the overflow of raw code when rendering a code block, the view of the end-user avoids the slider effect.

And, by eliminating the slider effect, responsive views using other devices ie.) Kindle Fire HDX will maintain screen boundaries without overflow.

Site Monitoring

Note. The Http Headers of this repo are being monitored by Internet Supervision dot com.

Reproduction In Part

Most authors will allow for the reproduction of their works in part ...

When the case of brief quotations embodied in critical articles or reviews is addressed.

As long as an appropriate citation is presented concurrent with the review, authors are generally receptive.

Therefore, please remember to provide a Link-back to the original author, or to the publisher of the original publication when citing.

Thank you for your consideration.

Copyright © 2000 - 2018

The Medical Marijuana Initiative of North America Small Registered MMINAIL™ Logo Badge Small Registered MMINAIL™ Logo Badge
- International Limited Small R-Reggae™ Trademark Symbol Logo Badge Small R-Reggae™ Trademark Symbol Logo Badge

This Open Source project is being hosted with Jekyll via GitHub Pages.

All rights reserved. Except where explicity granted.

Powered with glyphicons